Δ2,3 -1,4-Morpholine-2-carboxylic acid derivatives as antibacterial agents

ABSTRACT

There is described the stereoselective total synthesis by a variety of routes of novel suitably substituted Δ 2 ,3 -1,4-morpholine-2-carboxylic acids possessing a fused β-lactam ring in the 1,6-position and carrying a substituent cis to carbon 5 in the 7-position of the fused ring system represented by the general formula ##STR1## wherein Q is hydrogen, alkyl, aralkyl or --CH 2  COOZ where Z is hydrogen or the residue of an ester group and X is amino, azido or acylamino. Also included in the invention are compounds of the above formula in which the carboxyl group at the 2-position is protected as by an easily cleavable ester group and salts of both the free acids and carboxyl-protected compounds. Those compounds in which X is acylamino and their physiologically hydrolyzed esters and pharmaceutically acceptable salts are potent antibacterial agents.

CROSS-REFERENCE TO RELATED APPLICATION

This is a divisional application of co-pending application Ser. No.538,271 filed Jan. 2, 1975, now U.S. Pat. No. 4,012,383.

BACKGROUND OF THE INVENTION

(1) Field of the Invention

The chemical processes of the present invention produce novelantibacterial agents of the β-lactam type containing a hitherto unknownnucleus and useful intermediates for their synthesis.

(2) Description of the Prior Art

Penicillins and cephalosporins comprise a group of well-knownantibacterial agents commonly grouped together as a class calledβ-lactam antibiotics. For a recent review of this field with manycitations (especially the first ten) to the prior work, see J. P. Houand J. W. Poole, β-lactam Antibiotics: Their Physiocochemical Propertiesand Biological Activities in Relation to Structure, J. PharmaceuticalSciences, 60(4), 503-532. (April, 1971). Most of the work in this fieldhas fundamentally been done, speaking broadly, with 6-aminopenicillanicacid, 7-aminocephalosporanic acid and derivatives thereof produced byfermentation or chemical transformation of the natural products. Despitethe extensive progress made in preparing active derivatives of6-aminopenicillanic acid and 7-aminocephalosporanic acid, there is acontinuing search for synthetic and semi-synthetic routes to newfamilies of β-lactam antibiotics which may possess more advantageousproperties than those derived from the known pencillin and cephalosporinnuclei.

Considerable work has been done on total chemical synthesis of bothknown β-lactams and nuclear analogs of such known compounds. A recentreview is the text by M. S. Manhas and A. K. Bose, Synthesis ofPenicillin, Cephalosporin C and Analogues, Marcel Decker, Inc., 95Madison Avenue, New York, New York, 1969. Another extensive review is byR. B. Morin and B. G. Jackson, Chemistry of Cephalosporin Antibiotics,Fortschr. Chem. Org. Naturst., 28, 343-403 (1970), especially pages379-393; the now famous "Woodward Intermediate" is shown therein ascompound 146 on page 387. A more recent review of β-lactams is that byM. S. Manhas and A. K. Bose, Beta-Lactams: Natural and Synthetic: Part1, Wiley-Interscience, New York, New York, 1971. A still further reviewarticle on the synthesis of β-lactams is that by A. K. Mukerjee et al.,Synthesis, 327 (1973).

Other pertinent publications relating to synthesis of β-lactams are:

(a) D. M. Brunwin, G. Lowe and J. Parker J.C.S. Chem. Comm., 1971,865-867, describing synthesis of nuclear analogs of thepenicillin-cephalosporin group.

(b) D. M. Brunwin et al., J. Chem. Soc. (C), 1971, 3756-3762 and J.C.S.Chem. Comm., 1972, 589-590 describing total synthesis of nuclear analogsof penicillins and cephalosporins.

(c) S. Kukolja, J. Amer. Chem. Soc., 93, 6267-6270 (1971) and 94,7590-7593 (1972) describing preparation of6-phthalimido-5-epipenicillanates and disulfide analogs of penicillins.

(d) J. A. Webber et al., J. Medicinal Chemistry, 14(11), 1136-1138(1971) describing preparation of 3-cyanomethyl cephem nucleus.

(e) West German Patent Specification No. 2,219,601 (Farmdoc 76,051T)describing synthesis of β-lactams of the formula ##STR2## wherein X ishalo, N₃ -- or H₂ N--, A is --S--, --S--CH₂ --, --O--, --O--CH₂ --,--CH₂, --CH₂ CH₂ -- or --NH-- and R¹, R² and R³ are hydrogen, C₁ --C₆alkyl or aryl.

(f) U.K. Pat. No. 1,308,822 disclosing β-lactams of the formula ##STR3##where Y = amino or substituted amino.

(g) S. Wolfe et al., Can. J. Chem., 50, 2894-2905 (1972) describingsynthesis of sulfur-free penicillin derivatives.

(h) French Pat. No. 2,111,859 describing nuclei of the formula ##STR4##and 7-acylated derivatives thereof.

(i) Helvetica Chimica Acta, 55(2), 388-429 (1972) describing nuclearmodified cephalosporins and penicillins.

(j) F. Moll et al., Zeit. fur Naturforsch. B, 27(b)6, 727 (1972)describing nuclear analogs of cephalosporins.

(k) U.K. Specification Nos. 1,271,013 and 1,271,014 describing γ-lactamsof 7-(acylamino)-3-aminomethyl-ceph-3-em-4-carboxylic acids.

(1) U.K. Specification No. 1,271,180 describing preparation of novelthiazoline azetidinone rearrangement products useful as intermediates inpenicillin and cephalosporin synthesis.

(m) German Patent Specification Nos. 2,046,822, 2,046,823 and 2,046,824describing synthesis of novel azetidinone intermediates.

(n) G. Lowe et al., J. Chem. Soc. Perkins I, 1322 (1973) describingtotal synthesis of nuclear analogs of 7-methylcephalosporins having theformula ##STR5##

(o) D. M. Brunwin et al., J. Chem. Soc. Chem. Comm., 865 (1971)describing synthesis of compounds of the formula ##STR6##

(p) S. Wolfe et al., Canadian J. Chem., 50, 2902 (1972) describingcompounds of the formula ##STR7##

(q) J. P. Luttringer et al., Tetrahedron Letters, 4163-4166 (1973)describing compounds of the formula ##STR8##

SUMMARY OF THE INVENTION

The present invention provides stereoselective total synthesis ofcertain novel substituted Δ²,3 -1,4-morpholine-2-carboxylic acidspossessing a fused β-lactam ring in the 1,6-position and carrying asubstituent cis to carbon 5 in the 7-position of the fused ring systemrepresented by the general formula ##STR9## wherein Q is hydrogen,alkyl, aralkyl or --CH₂ COOZ where Z is hydrogen or the residue of anester group and X is azido, amino or acylamino. When X is acylamino,these acids (and their pharmaceutically acceptable salts andphysiologically hydrolyzed esters) are potent antibacterial agents.

Also included in this invention are various novel intermediates usefulin preparing the active β-lactam derivatives described above and variousprocesses for the production of the intermediates and active compounds.

The compounds having the above general formula represent a new family ofβ-lactam antibiotics. They can be considered nuclear analogs ofcephalosporins in which the sulfur atom of the dihydrothiazine ring isreplaced by an oxygen atom and shifted from position 5 to 4 of theβ-lactam ring system as numbered in the formula above. The nomenclatureto be used could be the following: ##STR10##3-oxa-6-aza-bicyclo-[4,2,0]octan-7-one.

However, Sheehan has used the term O-cepham for the structure ##STR11##[J. C. Sheehan and M. Dadic, J. Heterocyclic Chem., 5, 770 (1968)] andwe propose the use of the term O-2-isocepham for the basic system havingthe formula ##STR12## The numerical prefix indicates the position of thehetero-atom.

To illustrate the above system, the intermediate of the formula##STR13## may be named benzyl 7β-amino-3-methyl-Δ³-O-2-isocephem-4-carboxylate and the compound of the formula ##STR14##named 7β-(2-aminomethylphenylacetamido)-3-methyl-Δ³-O-2-isocephem-4-carboxylic acid.

The general methods of synthesis for the compounds of the presentinvention are illustrated below for the case in which Q is methyl inCharts A & B, for Q as benzyl in Chart C, for Q as phenethyl in Chart D,for Q as --CH₂ COOZ in Chart E and for Q as hydrogen in Charts F, G, H,and J. Additional routes to desirable intermediates and finalantibacterial agents are to be found in Chart I and in the examples andother disclosure which follow. ##STR15##

There is thus provided by the present invention the novel O-2-isocephemcompounds having the formula ##STR16## wherein R is an acyl group and Qis hydrogen, C₁ -C₁₀ alkyl, aralkyl (preferably benzyl or phenethyl) or--CH₂ COOZ in which Z is hydrogen or the residue of an ester group(preferably C₁ -C₆ alkyl), and easily cleavable esters andpharmaceutically acceptable salts thereof.

The acyl group R can be chosen from a wide variety of organic acylradicals which yield products of improved properties and is preferablyan acyl radical which is contained in a naturally occurring orbio-synthetically, semi-synthetically or totally-synthetically activeN-acyl derivative of 6-aminopenicillanic acid or 7-aminocephalosporanicacid. Examples of suitable acyl groups are defined in the followinggeneral formulae, but it should be noted that this is not intended to bean exhaustive list of all the possible acyl groups which may be used.

    R.sup.a C.sub.n H.sub.2n CO--                              (i)

where R^(a) is aryl (carbocyclic or heterocyclic), substituted aryl,cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl or a nonaromatic or mesoionic heterocyclic group, and n isan integer from 1-4. The preferred R^(a) substituents are (a) arylselected from phenyl, 2-thienyl, 3-thienyl, furyl, 4-isoxazolyl,pyridyl, tetrazolyl, sydnone-3 or -4, imidazolyl, naphthoyl,quinoxalinyl, triazolyl, isothiazolyl, thiadiazolyl, thiazolyl,oxazolyl, oxadiazolyl, pyrazolyl, furazan, pyrazinyl, pyrimidinyl,pyridazinyl or triazinyl; (b) substituted aryl in which the aryl groupsmentioned above under (a) are substituted by one or more radicalsselected from chloro, bromo, iodo, fluoro, nitro, amino, cyano,(lower)alkanoyloxy, (lower)-alkanoyl, (lower)alkoxyamino, (lower)alkoxy,(lower)-alkyl, (lower)alkylamino, hydroxy, guanidino, (lower)-alkylthio,carboxy, phenyl, halophenyl, trifluoromethyl, di(lower)alkylamino,sulfamyl, (lower)alkanoylamino, phenyl(lower)alkylamido, cyclokylamino,allylamido morpholinocarbonyl, pyrrolidinocarbonyl, piperidinocarbonyl,tetrahydropyridino, furfurylamindo or N-alkyl-N-anilino; (c) C₃ -C₁₂cycloalkyl; (d) substituted C₃ -C₁₂ cycloalkyl where the substituentsare one or more radicals selected from chloro, bromo, fluoro, iodo,nitro, trifluoromethyl, C₁ -C₄ alkyl, C₁ -C₄ alkylamino, C₁ -C₂ alkoxyor amino; (e) C₃ -C₁₂ cycloalkenyl, said cycloalkenyl group having 1 or2 double bonds; and (f) substituted C₃ -C₁₂ cycloalkenyl, saidcycloalkenyl group having 1 or 2 double bonds and being substituted byone or more radicals selected from chloro, bromo, fluoro, iodo, nitro,trifluoromethyl, C₁ -C₄ alkyl, C₁ -C₄ alkylamino, C₁ -C₂ alkoxy oramino. The most preferred R^(a) groups are phenyl; phenyl substituted byone or more radicals selected from chloro, bromo, iodo, fluoro, nitro,amino, (lower)alkyl, guanidino, (lower)alkylthio, cyano, (lower)alkoxy,sulfamyl, (lower)alkylamino, hydroxy, acetoxy, or trifluoromethyl;2-thienyl; 3-thienyl; tetrazolyl; sydnone -3; sydnone -4; furyl;isothiazolyl; thiadiazolyl optionally substituted with phenyl;oxadiazolyl optionally substituted with phenyl; thiazolyl; imidazolyl;triazolyl; oxazolyl; pyridyl; furazan optionally substituted at the3-position with methoxy; 4-isoxazolyl optionally substituted at the5-position with methyl and at the 3-position with phenyl or halophenyl;1,4-cyclohexadienyl; 1-cyclohexenyl and 1-aminocyclohexyl.

The most preferred acyl groups of this category are those in which nis 1. Examples of this category include phenylacetyl, halophenylacetyl,nitrophenylacetyl, aminophenylacetyl, β-(o-aminomethylphenyl)propionyl,(lower)alkanoyloxyphenylacetyl (e.g. p-acetoxyphenylacetyl),(lower)alkoxyphenylacetyl (e.g. methoxyhenylacetyl, ethoxyphenylacetyl),(lower)alkylphenylacetyl (e.g. methylphenylacetyl or ethylphenylacetyl),hydroxyphenylacetyl (e.g. o-hydroxyphenylacetyl),(lower)alkylaminophenylacetyl (e.g. o-, m- or p-aminomethylphenylacetyl), o-m- or p- guanidinophenylacetyl,o-carboxyphenylacetyl, N,N-bis-(2-chloroethyl)aminophenylpropionyl,thien-2 and 3-ylacetyl, 2- or 3-furylacetyl, 1,2,5-thiadiazole-3-acetyl,isothiazolyl-4-acetyl, 4-isoxazolylacetyl, 1-cyclohexenylacetyl,2-aminomethyl-1-cyclohexenylacetyl, 1-aminocyclohexylacetyl,1,4-cyclohexadienylacetyl, 2-aminomethyl-1,4-cyclohexadienylacetyl,pyridylacetyl, tetrazolylacetyl (other heterocyclic groups of this typeare disclosed in U.S. Pat. Nos. 3,819,623 and 3,516,997) or asydnoneacetyl group as disclosed in U.S. Pat. Nos. 3,681,328, 3,530,123and 3,563,983. Other groups of this type include3-phenyl-5-chlorophenyl-5-methylisoxazol-4-ylacetyl and3-(2,6-dichlorophenyl)-5-methylisoxazol-4-ylacetyl or a group in whichisoxazolyl is replaced by isothiazole as disclosed in U.S. Pat. No.3,551,440. Still other examples are o-, m- andp-(2'-aminoethoxy)phenylacetyl (as disclosed in U.S. Pat. No.3,759,905), 4,5-dimethoxycarbonyl-1,2,3-triazol-1-ylacetyl or4-cyano-1,2,3-triazol-1-yl acetyl (as disclosed in U.S. Pat. No.3,821,206) and imidazol-(1)-acetyl (as disclosed in U.S. Pat. No.3,632,810;

    C.sub.n H.sub.2n+1 CO--                                    (ii)

where n is an integer from 1-7. The alkyl group may be straight orbranched and, if desired, may be interrupted by an oxygen or sulphuratom or substituted by, e.g., a cyano group. Examples of this groupinclude cyanoacetyl, valeryl, hexanoyl, heptanoyl, ethoxycarbonyl,octanoyl and butylthioacetyl. A preferred acyl group is cyanoacetyl;

    C.sub.n H.sub.2n-1 CO--                                    (iii)

where n is an integer from 2-7. The alkenyl group may be straight orbranched and, if desired, may be interrupted by an oxygen or sulphuratom. An example of this group is allylthioacetyl; ##STR17## where R^(a)is as defined under (i) and in addition may be benzyl, C₁ -C₆ alkyl or(lower)alkoxycarbonyl and R^(b) and R^(c) which may be the same ordifferent each represent hydrogen, phenyl, benzyl, phenethyl or C₁ -C₆alkyl. The preferred R^(a) substituents in this category are benzyl, C₁-C₆ alkyl, (lower)alkoxycarbonyl and those mentioned under (i) as beingpreferred aryl, substituted aryl, cycloalkyl (and substitutedcycloalkyl) and cycloalkenyl (and substituted cycloalkenyl) groups. Themost preferred R^(a) group is phenyl. Examples of this group includephenoxyacetyl, 2-phenoxy-2-phenylacetyl 2-phenoxypropionyl,2-phenoxybutyryl, benzyloxyacetyl, 2-methyl-2-phenoxypropionyl,p-cresoxyacetyl, p-methylthiophenoxyacetyl and ethoxycarbonylacetyl;##STR18## where R^(a) is as defined under (i) and in addition may bebenzyl or C₁ -C₆ alkyl and R^(b) and R^(c) have the meanings definedunder (iv). The preferred R^(a) substituents in this category arebenzyl, -C₁ -C₆ alkyl and those mentioned under (i) as being preferredaryl, substituted aryl, cycloalkyl (and substituted cycloalkyl) andcycloalkenyl (and substituted cycloalkenyl) groups. The most prefrredaryl groups of this type are those in which R^(b) and R^(c) are hydrogenand R^(a) is pheny; phenyl substituted with one or more radicalsselected from chloro, bromo, iodo, fluoro, nitro, amino, (lower)alkyl,(lower)alkythio, cyano, (lower)alkoxy, (lower)alkylamino, hydroxy,acetoxy or trifluoromethyl; 3-pyridyl; or 4-pyridyl;

    R.sup.a X (CH.sub.2).sub.m CO--                            (vi)

where R^(a) is as defined under (i) and in addition may be benzyl, X isoxygen or sulphur and m is an integer from 2-5. The preferred R^(a)groups are benzyl and those mentioned under (i) as being preferred aryl,substituted aryl, cycloalkyl (and substituted cycloalkyl) andcycloalkenyl (and substituted cycloalkenyl) groups. An example of thisgroup is S-benzylthiopropionyl.

    R.sup.a CO--                                               (vii)

where R^(a) is as defined under (i). The preferred R^(a) groups arethose mentioned under (i) as being preferred aryl, substituted aryl,cycloalkyl (and substituted cycloalkyl and cycloalkenyl (and substitutedcycloalkenyl) groups. The most preferred aryl groups of this categoryare those in which R^(a) is phenyl; phenyl substituted with one or moreradicals selected from chloro, bromo, iodo, fluoro, nitro, amino,(lower)alkyl, (lower)alkylthio, cyano, (lower)alkoxy, (lower)alkylamino,di(lower)alkylamino, hydroxy, acetoxy or trifluoromethyl, and mostpreferably phenyl substituted at the 2-position by carboxy or phenyl orat the 2- and 6-positions by methoxy; 2-ethoxynaphthoyl;3-phenyl-5-methylisoxazol-4-yl; 3-o-chlorophenyl-5-methylisoxazol-4-yl;3-(2,6-dichlorophenyl)-5-methylisoxazol-4-yl and 1-aminocyclohexyl.Examples of this group includes 2,6-dimethoxybenzoyl, benzoyl,2-biphenylcarbonyl, 2-aminomethylbenzoyl,2-carboxybenzoyl-2-phenylbenzoyl, 2-thienylcarbonyl, 3-thienylcarbonyland 2-chlorobenzoyl; ##STR19## where R^(a) is as defined under (i) and Yis hydrazino, guanidino, ureido, thioureido and substituted thioureido(as disclosed in U.S. Pat. No. 3,741,962), allophanamido (as describedin U.S. Pat. No. 3,483,188), 3-guanyl-1-ureido ( as in U.S. Pat. No.3,796,709), 3-(2-furoyl)ureido, cyanamino (as in U.S. Pat. No.3,796,709), 3-(benzoyl)ureido, azido, amino, acylamino (e.g.carbobenzoxyamino), a group obtained by reacting the amino group of the7-side chain with an aldehyde or ketone (e.g. acetone, formaldehyde,acetaldehyde, butyraldehyde, acetylacetone, methyl acetoacetate,benzaldehyde, salicylaldehyde, methyl ethyl ketone or ethylacetoacetate), hydroxy, etherified hydroxy, esterified hydroxy, carboxy,esterified carboxy (as disclosed for example in U.S. Pat. Nos.3,282,926, 3,819,601 and 3,635,961 and including especially ##STR20##tetrazolyl, cyano, halogeno, acyloxy (e.g. formyloxy or(lower)alkanoyloxy), sulfo, sulfoamino or esterified sulfo. Thepreferred R^(a) substituents are those mentioned under (i) as beingpreferred aryl, substituted aryl, cycloalkyl (and substitutedcycloalkyl) and cycloalkenyl (and substituted cycloalkenyl) groups.Preferred Y substituents are hydrazino; guanidino; ureido; substitutedthioureido of the formula ##STR21## in which R^(p) is hydrogen or C₁ -C₈alkyl and R^(q) is hydrogen, C₁ -C₈ alkyl, C₂ -C₈ alkenyl, phenyl,benzoyl, C₁ -C₈ alkoxy- C₁ -C₈ alkyl, (carbo-C₁ -C₈ alkoxy) C₁ -C₈alkyl; allophanamido; 3-guanyl-1-ureido; 3-(2-furoyl)ureido;3-(benzoyl)ureido; azido; amino; a group obtained by reacting the aminogroup Y with acetone, formaldehyde, acetaldehyde, butyraldehyde,acetylacetone, methyl acetoacetate, benzaldehyde, salicylaldehyde,methyl ethyl ketone or ethyl acetoacetate; hydroxy; etherified hydroxyincluding especially (lower)alkoxy; carboxy; esterified carboxyincluding especially 5-indanyloxycarbonyl; triazolyl; tetrazolyl; cyano;cyanamino; halogeno; formyloxy; (lower)alkanoyloxy; sulfo; orsulfoamino. Examples of this group include α-aminophenylacetyl;α-carboxyphenylacetyl; 2,2-dimethyl-5-oxo-4-phenyl-1-imidazolyl;α-amino-p-hydroxyphenylacetyl; α-hydroxyphenylacetylα-formyloxyphenylacetyl and other aryl groups of this type disclosed inU.S. Pat. Nos. 3,812,116 and 3,821,017; α-amino-α-2- or 3-thienylacetyl;α-amino-60-(3-chloro-4-hydroxy)phenylacetyl;α-amino-α-(1,4-cyclohexadienyl)acetyl; α-azidophenylacetyl; α-amino-α-(1-cyclohexenyl)acetyl; 2-carboxy-α-3-thienylacetyl;α-amino-α-(3,5-dichloro-4-hydroxyphenyl)acetyl; α-amino-α-3- or 4- or5-isothiazolylacetyl (as in U.S. Pat. No. 3,579,506) and other α-aminoand α-hydroxy-heterocyclylacetyl groups as disclosed for example in U.S.Pat. No. 3,821,207; ##STR22## where R^(d), R^(e) and R^(f) which may bethe same or different may each represent C₁ -C₆ alkyl, phenyl orsubstituted phenyl. The preferred phenyl substituents are one or moreradicals selected from chloro, bromo, iodo, fluoro, trifluoromethyl,nitro, amino, cyano, (lower)alkanoyloxy, (lower)alkanoyl,(lower)alkoxyamino, (lower)alkoxy, (lower)alkyl, (lower)alkylamino,hydroxy, (lower)alkylthio, carboxy, di(lower)alkylamino or sulfamyl. Anexample of this group is triphenylmethylcarbonyl. ##STR23## where R^(a)is as defined under (i) and in addition may be hydrogen, C₁ -C₆ alkyl,halogen substituted C₁ -C₆ alkyl, phenethyl, phenoxymethyl; benzyl or##STR24## and X is oxygen or sulphur. An example of such a group isCl(CH₂)₂ NHCO; ##STR25## where Y is as defined under (viii) and n is aninteger of 1-4. A most preferred Y substituent is amino. An example ofthis group is 1-aminocyclohexanecarbonyl.

Aminoacyl, for example

    R.sup.g CH(NH.sub.2)--(CH.sub.2).sub.n CO--                (xii)

where n is an integer of 1-10, or

    H.sub.2 N--C.sub.n H.sub.2n Ar(CH.sub.2).sub.m CO--

where m is zero or an integer from 1-10, and n is 0, 1, or 2; R^(g) ishydrogen or an alkyl, aryl, aralkyl or carboxy group or a group asdefined under R^(a) in (i) above; and Ar is an arylene group, e.g.p-phenylene or 1,4-naphthylene. Preferred aryl groups of the aboveformulae are those in which R^(g) is hydrogen, (lower)alkyl, phenyl,benzyl or carboxy and Ar is p-phenylene or 1,4-naphthylene. Examples ofsuch groups are disclosed in U.K. Pat. No. 1,054,806. Examples of groupsof this type include p-aminophenylacetyl and δ-aminoadipoyl derived fromnaturally occurring amino acids and derivatives thereof, e.g.N-benzoyl-δ-aminoadipoyl;

Substituted glyoxylyl groups of the formula

    R.sup.h.CO.CO--                                            (xiii)

where R^(h) is an aliphatic, araliphatic or aromatic group. Thepreferred R^(h) groups are 2-thienyl; 3-thienyl; α-naphthyl;2-phenanthryl or a mono-, di- or tri-substituted phenyl group, thesubstituents being selected from chloro, bromo, iodo, fluoro, amino,di(lower)alkylamino, (lower)alkyl, (lower)alkoxy, nitro or(lower)alkanoylamino. Examples of this category are disclosed in U.S.Pat. Nos. 3,546,219 and 3,573,294. Included in this group are also theα-carbonyl derivatives of the above substituted glyoxylyl groups formedfor example with hydroxylamine, semicarbazide, thiosemicarbazide,isoniazide or hydrazine; ##STR26## where R^(a) has the meaning definedunder (i), X is oxygen or sulphur, X' is oxygen or imino and R¹represents (lower)alkyl, cycloalkyl having 4, 5, 6 or 7 carbon atoms,monohalo(lower)alkyl, dichloromethyl, trichloromethyl, (lower)alkenyl of2-6 carbon atoms, ##STR27## n is an integer from 0 to 3 inclusive andeach of R^(k) and R^(j) is hydrogen, nitro, di(lower) alkylamino,(lower)-alkanoylamino, (lower)alkanoyloxy, C₁ -C₆ alkyl, C₁ -C₆ alkoxy,sulfamyl, chloro, bromo, iodo, fluoro or trifluoromethyl. The preferredR^(a) substituents are those mentioned under (i) as being preferredaryl, substituted aryl, cycloalkyl (and substituted cycloalkyl) andcycloalkenyl (and substituted cycloalkenyl) groups. Preferred acylgroups of this type are those in which R^(a) is 2-thienyl; 3-thienyl;phenyl; or phenyl substituted by one or more radicals selected fromnitro, di(lower)alkylamino, (lower)alkanoylamino, amino, hydroxy,(lower)alkanoyloxy, C₁ -C₆ alkyl, C₁ -C₆ alkoxy, sulfamyl, chloro,bromo, iodo, fluoro or trifluoromethyl; X is oxygen; X' is oxygen orimino and R^(i) is (lower)alkyl, phenyl, 2-thienyl, 3-thienyl, 2-furylor 5-nitro-2-furyl. The most preferred groups are those of the aboveformula where R^(a) is phenyl, p-hydroxyphenyl, 2-thienyl or 3-thienyl;X is oxygen; X' is oxygen, and R^(i) is phenyl or 2-furyl. Examples aredisclosed in U.S. Pat. Nos. 3,687,949 and 3,646,024; ##STR28## whereR^(a) has the meaning defined in (i) and R^(i) has the meaning definedin (xiv). The preferred R^(a) substituents are those mentioned under (i)as being preferred aryl, substituted aryl, cycloalkyl (and substitutedcycloalkyl) and cycloalkenyl (and substituted cycloalkenyl) groups.Preferred R^(i) substituents include (lower)alkyl, dichloromethyl, C₄-C₇ cycloalkyl, 2-thienyl, 3-thienyl, phenyl, benzyl, halobenzyl,##STR29## Examples of this group are disclosed in U.S. Pat. Nos.3,626,024 and 3,692,779; ##STR30## where R^(a) has the meaning definedin (i) and R¹ is (lower)alkyl, C₃ -C₁₂ cycloalkyl, aryl (especiallyphenyl), a monocyclic heterocylcic radical having 5 or 6 atoms exclusiveof hydrogen which are C, S, N or O, no more than 2 atoms being otherthan C, or a substituted monocyclic heterocyclic radical as definedabove having one or more substituents selected from halo, (lower)alkyl,(lower)alkoxy or phenyl. Examples of this group are disclosed in U.S.Pat. No. 3,778,436. Most preferred R¹ groups are (lower)alkyl, phenyl,thienyl or furyl.

A preferred class of acyl groups are those of the formula ##STR31##wherein Ar' is a radical of the formula ##STR32## in which R^(m), R^(n)and R^(o) are alike or different and each is hydrogen, hydroxy,(lower)alkyl, cyano, (lower)alkoxy, chloro, bromo, iodo, fluoro,trifloromethyl, nitro, amino, (lower)alkylamino, di(lower)alkylamino,(lower)alkanoyl, (lower)alkanoyloxy such as p-acetoxy or phenyl and Y isamino or a group obtained by reacting the amino group with acetaldehyde,formaldehyde or acetone, fluoro, chloro, bromo, iodo, hydroxy,(lower)alkanoyloxy, carboxy, guanidino, 3-guanyl-1-ureido,3-(2-furoyl)ureido, 3-benzoylureido, sulfo, sulfoamino, ureido,thioureido, (lower)alkoxy, cyano, cyanamino or indanyloxycarbonyl.Particularly preferred Ar radicals are phenyl, p-hydroxyphenyl,4-hydroxy-3,5-dichlorophenyl, 3-chloro-4-hydroxyphenyl, o-, m- or p-aminomethylphenyl, 2-thienyl, 3-thienyl, 1-cyclohexenyl and1,4-cyclohexadienyl. Particularly preferred Y groups are amino, hydroxyand carboxy. Set forth below are formulae of the most preferred acylgroups of this class: ##STR33## Of most interest are the acyl groups ofthe above class where the acid ArCH(X)COOH is of the D-series.

Other particularly preferred acyl groups for the compounds of formula Iare ##STR34## where U and V are alike or different and each is hydrogen,chloro or fluoro; ##STR35##

The term "(lower)alkyl" as used herein means both straight and branchedchain aliphatic hydrocarbon radicals having from one to ten carbon atomssuch as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl,amyl, hexyl, 2-ethylhexyl, heptyl, decyl, etc. Similarly, where the term"(lower)" is used as part of the description of another group, e.g."(lower)alkoxy", it refers to the alkyl portion of such group which istherefore described above in connection with "(lower)alkyl".

The pharmaceutically acceptable salts referred to above include thenontoxic carboxylic acid salts e.g. nontoxic metallic salts such assodium, potassium, calcium and aluminum, the ammonium salt and saltswith nontoxic amines, e.g., trialkyleneamines, procaine, dibenzylamine,N-benzyl-β-phenethylamine, 1-ephenamine, N,N'-dibenzylethylenediamine,N-alkylpiperidine and other amines which have been used to form salts ofpenicillins and cephalosporins. When a basic group is present, as whenit occurs in the 7-acyl group, the present invention also includes thepharmaceutically acceptable acid addition salts, e.g. salts with mineralacids such as hydrochloric, hydrobromic, hydroiodic, phosphoric,sulfuric and salts with organic acids such as maleic, acetic, citric,oxalic, succinic, benzoic, tartaric, fumaric, mandelic, ascorbic andmalic. The term "pharmaceutically acceptable salts" is also meant toinclude nontoxic acid addition salts of the easily cleavable estersreferred to above. The compounds which contain a basic group in radicalR may also be present in the form of an internal salt, i.e. in the formof the zwitterion.

The easily cleavable esters referred to above include ester groups whichare removable by methods, e.g. chemical or enzymatic hydrolysis,treatment with chemical reducing agents under mild conditions,irradiation with ultraviolet light or catalytic hydrogenation, which donot result in any appreciable destruction of the remaining portion ofthe molecule. Examples of suitable esters include those disclosed inU.S. Pat. Nos. 3,284,451 and 3,249,622 and U.K. Pat. Nos. 1,229,453 and1,073,530. Esters which have been used previously in penicillin andcephalosporin chemistry include for example benzhydryl, p-nitrobenzyl,benzyl, trichloroethyl, silyl such as trimethylsilyl, phenacyl,p-methoxybenzyl, acetonyl, phthalidyl, indanyl and (lower)alkyl such asmethyl, ethyl and t-butyl. Particularly preferred easily cleavableesters are those which are hydrolyzed under physiological conditionssuch as pivaloyloxymethyl, acetoxymethyl, phthalidyl, indanyl andmethoxymethyl.

As the O-2-isocephem compounds of the present invention may possess oneor more asymmetric carbon atoms, the invention includes all of thepossible enantiomeric and diastereomeric forms of the compounds of thegeneral formula I shown above. Resulting mixtures of isomers can beseparated into the individual isomers according to methods which areknown per se, e.g. fractional crystallization, adsorption chromatographyor other suitable separation processes. Resulting racemates can beseparated into the antipodes in the usual manner after introduction ofsuitable salt-forming groupings, e.g. by forming a mixture ofdiastereoisomeric salts with optically active salt-forming agents,separating the mixture into diastereoisomeric salts, and converting theseparated salts into the free compounds, or by fractionalcrystallization from optically active solvents.

It will be appreciated that certain of the compounds of this inventionexist in various states of solvation and the anhydrous as well assolvated forms are within the scope of the invention.

The free acid compounds of the above general formula I where R is acyland physiologically hydrolyzed esters thereof together with thepharmaceutically acceptable salts of such free acids and esters areactive antibacterial agents. The remaining compounds of the abovegeneral formula I including salts thereof are valuable intermediateswhich can be converted into the above-mentioned pharmacologically activecompounds in a simple manner, for example, as described below.

Preferred embodiments of the present invention are the O-2-isocephemcompounds having the formula ##STR36## wherein R represents an acylgroup, and easily cleavable esters and pharmaceutically acceptable saltsthereof.

Preferred acids, esters and salts of formula II are those in which acylgroup R is selected from the acyl groups defined above under (i) to(xvi). The acyl groups mentioned as being preferred groups withincategories (i) to (xvi) are also preferred in the compounds defined bygeneral formula II.

More preferred acids, esters and salts of formula II are those in whichacyl group R is ##STR37## in which Ar' is a radical of the formula##STR38## in which R^(m), R^(n) and R^(o) are alike or different andeach is hydrogen, hydroxy, (lower)alkyl, cyano, (lower)alkoxy, chloro,bromo, iodo, fluoro, trifluoromethyl, nitro, amino, (lower)alkylamino,di(lower)alkylamino, (lower)alkanoyl, (lower)alkanoyloxy such asp-acetoxy or phenyl and Y is amino or a group obtained by reacting theamino group with acetaldehyde, formaldehyde or acetone, fluoro, chloro,bromo, iodo, hydroxy, (lower)alkanoyloxy, carboxy, guanidino,3-guanyl-1-ureido, 3-(2-furoyl)ureido, 3-benzoylureido, sulfo,sulfoamino, ureido, thioureido, (lower)alkoxy, cyano, cyanamino orindanyloxycarbonyl.

Other preferred acids, esters and salts of formula II are those in whichR is ##STR39## wherein Ar' is phenyl, p-hydroxyphenyl,4-hydroxy-3,5-dichlorophenyl, 3-chloro-4-hydroxyphenyl, o-, m- orp-aminomethylphenyl, 2-thienyl, 3-thienyl, 1-cyclohexenyl or1,4-cyclohexadienyl and Y is amino, carboxy or hydroxy.

Other preferred compounds of formula II are those wherein R is an acylgroup of the formula ##STR40##

Other preferred compounds of formula II are those wherein R is an acylgroup of the formula ##STR41## wherein U and V are alike or differentand each is hydrogen, chloro or fluoro; ##STR42##

A most preferred group of compounds are those acids defined by formulaII wherein R is α-carboxyphenylacetyl, cyanoacetyl,α-amino-α-(p-hydroxyphenyl)acetyl,α-amino-α-(3-chloro-4-hydroxyphenyl)-acetyl,α-amino-α-(3,5-dichloro-4-hydroxyphenyl)acetyl,α-amino-α-(2-thienyl)acetyl, α-amino-α-(3-thienyl)acetyl,α-amino-α-(1-cyclohexenyl)acetyl, α-amino-α-(1,4-cyclohexadienyl)acetyl,α-hydroxyacetyl, α-hydroxy-α-(2-thienyl)acetyl,α-hydroxy-α-(3-thienyl)acetyl, α-hydroxy-α-(1-cyclohexenyl)acetyl,α-hydroxy-α-(1,4-cyclohexadienyl)acetyl, α-carboxy-α-(2-thienyl)acetyl,α-carboxy-α-(3-thienyl)acetyl, α-carboxy-α-(1-cyclohexenyl)acetyl,α-carboxy-α-(1,4-cyclohexadienyl)acetyl,α-indanyloxycarbonyl-α-phenylacetyl, 1-(1H)-tetrazolyl,4-pyridylthioacetyl, 2-thienylacetyl, 3-thienylacetyl,1-cyclohexenylacetyl, 1,4-cyclohexadienylacetyl,o-aminomethylphenylacetyl, 1-aminocyclohexylcarbonyl,2,6-dimethoxybenzoyl, sydnoneacetyl or α-azidophenylacetyl, or apharmaceutically acceptable salt thereof.

Another most preferred group of compounds are the D-isomers of thoseacids defined by formula II wherein R isα-amino-α-(p-hydroxyphenyl)acetyl,α-amino-α-(3-chloro-4-hydroxyphenyl)acetyl,α-amino-α-(3,5-dichloro-4-hydroxyphenyl)acetyl,α-amino-α-(2-thienyl)acetyl, α-amino-α-(3-thienyl)acetyl,α-amino-α-(1-cyclohexenyl)acetyl, α-amino-α-(1,4-cyclohexadienyl)acetyl,α-hydroxyacetyl, α-hydroxy-α-(2-thienyl)acetyl, α-hydroxy-α-(3-thienyl)acetyl, α-hydroxy-α-(1-cyclohexenyl)acetyl orα-hydroxy-α-(1,4-cyclohexadienyl)acetyl, or a pharmaceuticallyacceptable salt thereof.

A most preferred compound of formula II is the acid in which R isphenoxyacetyl, or a pharmaceutically acceptable salt thereof.

Another most preferred compound of formula II is the acid in which R isphenylacetyl, or a pharmaceutically acceptable salt thereof.

A still further most preferred compound of formula II is the acid inwhich R is α-aminophenylacetyl, or a pharmaceutically acceptable saltthereof. The isomer of this compound in which the α-carbon atom of the7-acyl group is of the D-series is of particular importance due to itscombination of good antibacterial activity and oral absorption.

Other preferred embodiments of the present invention are theO-2-isocephem compounds having the general formula ##STR43## wherein Rrepresents an acyl group and W is C₁ -C₁₀ alkyl or aralkyl, (preferablybenzyl or phenethyl), and easily cleavable esters and pharmaceuticallyacceptable salts thereof.

Preferred acids, esters and salts of formula III are those in which acylgroup R is selected from the acyl groups defined above under (i) to(xvi). The acyl groups mentioned as being preferred groups withincategories (i) to (xvi) are also preferred in the compounds defined bygeneral formula III.

More preferred acids, esters and salts of formula III are those in whichacyl group R is ##STR44## in which Ar' is a radical of the formula##STR45## in which R^(m), R^(n) and R^(o) are alike or different andeach is hydrogen, hydroxy, (lower)alkyl, cyano, (lower)alkoxy, chloro,bromo, iodo, fluoro, trifluoromethyl, nitro, amino, (lower)alkylamino,di(lower)alkylamino, (lower)alkanoyl, (lower)alkanoyloxy such asp-acetoxy or phenyl and Y is amino or a group obtained by reacting theamino group with acetaldehyde, formaldehyde or acetone; fluoro; chloro;bromo; iodo; hydroxy; (lower)alkanoyloxy; carboxy; guanidino;3-guanyl-1-ureido; 3-(2-furoyl) ureido; 3-benzoylureido; sulfo;sulfoamino; ureido; thioureido; (lower)alkoxy; cyano; cyanamino; orindanyloxycarbonyl.

Other preferred acids, esters and salts of formula III are those inwhich R is ##STR46## wherein Ar' is phenyl, p-hydroxyphenyl,4-hydroxy-3,5-dichlorophenyl, 3-chloro-4-hydroxyphenyl, o-, m- orp-aminomethylphenyl, 2-thienyl, 3-thienyl, 1-cyclohexenyl or1,4-cyclohexadienyl and Y is amino, carboxy or hydroxy.

Other preferred compounds of formula III are those wherein R is an acylgroup of the formula ##STR47##

Other preferred compounds of formula III are those wherein R is an acylgroup of the formula ##STR48## wherein U and V are alike or differentand each is hydrogen, chloro or fluoro; ##STR49##

A most preferred group of compounds are those acids defined by formulaIII wherein R is α-carboxyphenylacetyl, cyanoacetyl,α-amino-α-(p-hydroxyphenyl)acetyl,α-amino-α-(3-chloro-4-hydroxyphenyl)acetyl,α-amino-α-(3,5-dichloro-4-hydroxyphenyl)acetyl,α-amino-α-(2-thienyl)acetyl, α-amino-α-(3-thienyl)acetyl,α-amino-α-(1-cyclohexenyl)acetyl, α-amino-α-(1,4-cyclohexadienyl)acetyl,α-hydroxyacetyl, α-hydroxy-α-(2-thienyl)acetyl,α-hydroxy-α-(3-thienyl)acetyl, α-hydroxy-α-(1-cyclohexenyl)acetyl,α-hydroxy-α-(1,4-cyclohexadienyl)acetyl, α-carboxy-α-(2-thienyl)acetyl,α-carboxy-α-(3-thienyl)acetyl, α-carboxy-α-(1-cyclohexenyl)acetyl,α-carboxy-α-(1,4-cyclohexadienyl)acetyl,α-indanyloxycarbonyl-α-phenylacetyl, 1-(1H)-tetrazolyl,4-pyridylthioacetyl, 2-thienylacetyl, 3-thienylacetyl,1-cyclohexenylacetyl, 1,4-cyclohexadienylacetyl,o-aminomethylphenylacetyl, 1-aminocyclohexylcarbonyl,2,6-dimethoxybenzoyl, sydnoneacetyl or α-azidophenylacetyl, or apharmaceutically acceptable salt thereof.

Another most preferred group of compounds are the D-isomers of thoseacids defined by formula III wherein R isα-amino-α-(p-hydroxyphenyl)acetyl,α-amino-α-(3-chloro-4-hydroxyphenyl)acetyl,α-amino-α-(3,5-dichloro-4-hydroxyphenyl)acetyl,α-amino-α-(2-thienyl)acetyl, α-amino-α-(3-thienyl)acetyl,α-amino-α-(1-cyclohexenyl)acetyl, α-amino-α-(1,4-cyclohexadienyl)acetyl,α-hydroxyacetyl, α-hydroxy-α-(2-thienyl)acetyl,α-hydroxy-α-(3-thienyl)acetyl, α-hydroxy-α-(1-cyclohexenyl)acetyl orα-hydroxy-α-(1,4-cyclohexadienyl)acetyl, or a pharmaceuticallyacceptable salt thereof.

A most preferred group of compounds of formula III are the acids inwhich R is phenoxyacetyl, or a pharmaceutically acceptable salt thereof.

Another most preferred group of compounds of formula III are the acidsin which R is phenylacetyl, or a pharmaceutically acceptable saltthereof.

A still further most preferred group of compounds of formula III are theacids in which R is α-aminophenylacetyl, or a pharmaceuticallyacceptable salt thereof. The isomers of these compounds in which theα-carbon atom of the 7-acyl group is of the D-series are of particularimportance due to their combination of good antibacterial activity andoral absorption.

Substituent W in the compounds of formula III above may be a C₁ -C₁₀alkyl or aralkyl radical. Thus, the alkyl radical may be any saturatedstraight, branched or cyclic monovalent hydrocarbon radical having from1 to 10 carbon atoms. The aralkyl substituent may be any substituent inwhich a mono-, bi- or polycyclic aromatic radical, said radical beingeither carbocyclic or heterocyclic and being optionally mono-, di- orpolysubstituted, is substituted for a hydrogen atom of an alkyl radical,said alkyl radical preferably being a saturated straight, branched orcyclic monovalent hydrocarbon radical having from 1 to 10 carbon atoms.

The most preferred compounds of general formula III are those in which Wis methyl, benzyl or phenethyl.

A most preferred series of compounds of the present invention are theO-2-isocephem compounds having the general formula ##STR50## wherein Rrepresents an acyl group, and easily cleavable esters andpharmaceutically acceptable salts thereof.

Preferred acids, esters and salts of formula IIId are those in whichacyl group R is selected from the acyl groups defined above under (i) to(xvi). The acyl groups mentioned as being preferred groups withincategories (i) to (xvi) are also preferred in the compounds defined bygeneral formula IIId.

More preferred acids, esters and salts of formula IIId are those inwhich acyl group R is ##STR51## in which Ar' is a radical of the formula##STR52## in which R^(m), R^(n) and R^(o) are alike or different andeach is hydrogen, hydroxy, (lower)alkyl cyano, (lower)alkoxy, chloro,bromo, iodo, fluoro, trifluoromethyl, nitro, amino, (lower)alkylamino,di(lower)alkylamino, (lower)alkanoyl, (lower)alkanoyloxy such asp-acetoxy or phenyl and Y is amino or a group obtained by reacting theamino group with acetaldehyde, formaldehyde or acetone; fluoro; chloro;bromo; iodo; hydroxy; (lower)alkanoyloxy; carboxy; guanidino;3-guanyl-1-ureido; 3-(2-furoyl)ureido; 3-benzoylureido; sulfo;sulfoamino; ureido; thioureido; (lower)alkoxy; cyano, cyanamino; orindanyloxycarbonyl.

Other preferred acids, esters and salts of formula IIId are those inwhich R is ##STR53## wherein Ar' is phenyl, p-hydroxyphenyl,4-hydroxy-3,5-dichlorophenyl, 3-chloro-4-hydroxyphenyl, o-, m- orp-aminomethylphenyl, 2-thienyl, 3-thienyl, 1-cyclohexenyl or1,4-cyclohexadienyl and Y is amino, carboxy or hydroxy. Other preferredcompounds of formula IIId are those wherein R is an acyl group of theformula ##STR54##

Other preferred compounds of formula IIId are those wherein R is an acylgroup of the formula ##STR55## wherein U and V are alike or differentand each is hydrogen, chloro or fluoro; ##STR56##

A most preferred group of compounds are those acids defined by formulaeIIId wherein R is α-carboxyphenylacetyl, cyanoacetyl,α-amino-α-(p-hydroxyphenyl)acetyl,α-amino-α-(3-chloro-4-hydroxyphenyl)acetyl,α-amino-α-(3,5-dichloro-4-hydroxyphenyl)acetyl,α-amino-α-(2-thienyl)acetyl, α-amino-α-(3-thienyl)acetyl,α-amino-α-(1-cyclohexenyl)acetyl, α-amino-α-(1,4-cyclohexadienyl)acetyl,α-hydroxyacetyl, α-hydroxy-α-(2-thienyl)acetyl,α-hydroxy-α-(3-thienyl)acetyl, α-hydroxy-α-(1-cyclohexenyl)acetyl,α-hydroxy-α-(1,4-cyclohexadienyl)acetyl, α-carboxy-α-(2-thienyl)acetyl,α-carboxy-α-(3-thienyl)acetyl, α-carboxy-α-(1-cyclohexenyl)acetyl,α-carboxy-α-(1,4-cyclohexadienyl)acetyl,α-indanyloxycarbonyl-α-phenylacetyl, 1-(1H)-tetrazolyl,4-pyridylthioacetyl, 2-thienylacetyl, 3-thienylacetyl,1-cyclohexenylacetyl, 1,4-cyclohexadienylacetyl,o-aminomethylphenylacetyl, 1-aminocyclohexylcarbonyl,2,6-dimethoxybenzoyl, sydnoneacetyl or α-azidophenylacetyl, or apharmaceutically acceptable salt thereof. The most preferred easilycleavable esters of this group are the pivaloyloxymethyl, methoxymethyl,indanyl, phthalidyl and acetoxymethyl esters and pharmaceuticallyacceptable salts thereof.

Another most preferred group of compounds are the D-isomers of thoseacids defined by formula IIId wherein R isα-amino-α-(p-hydroxyphenyl)acetyl,α-amino-α-(3-chloro-4-hydroxyphenyl)acetyl,α-amino-α-(3,5-dichloro-4-hydroxyphenyl)acetyl,α-amino-α-(2-thienyl)acetyl, α-amino-α-(3-thienyl)acetyl,α-amino-α-(1-cyclohexenyl)acetyl, α-amino-α-(1,4-cyclohexadienyl)acetyl,α-hydroxyacetyl, α-hydroxy-α-(2-thienyl)acetyl,α-hydroxy-α-(3-thienyl)acetyl, α-hydroxy-α-(1-cyclohexenyl)acetyl orα-hydroxy-α-(1,4-cyclohexadienyl)acetyl, or a pharmaceuticallyacceptable salt thereof. The most preferred easily cleavable esters ofthis group are the pivaloyloxymethyl, methoxymethyl, indanyl, phthalidyland acetoxymethyl esters and pharmaceutically acceptable salts thereof.

A most preferred acid of formula IIId is that in which R isphenoxyacetyl, or a pharmaceutically acceptable salt thereof.

A most preferred acid of formula IIId is that in which R isphenylacetyl, or a pharmaceutically acceptable salt thereof.

A most preferred acid of formula IIId is that in which R isα-aminophenylacetyl, or a pharmaceutically acceptable salt thereof.

A most preferred compound of formula IIId is the pivaloyloxymethyl,acetoxymethyl, indanyl, phthalidyl or methoxymethyl ester of the acid inwhich R is α-aminophenylacetyl, or a pharmaceutically acceptable saltthereof.

A still more preferred compound of formula IIId is the D-isomer of anacid of formula IIId in which R is α-aminophenylacetyl, or apharmaceutically acceptable salt thereof. This compound is found to beespecially valuable in view of its combination of good antibacterialactivity and oral absorption. The pivaloyloxymethyl, acetoxymethyl,indanyl, phthalidyl and methoxymethyl esters of the above acid as wellas pharmaceutically acceptable salts thereof are also preferredcompounds of the present invention.

Other preferred embodiments of the present invention are theO-2-isocephem compounds having the general formula ##STR57## wherein Rrepresents an acyl group and Z is hydrogen or the residue of an estergroup (preferably C₁ -C₆ alkyl), and easily cleavable esters andpharmaceutically acceptable salts thereof. The free carboxylic acids,physiologically hydrolyzed esters and pharmaceutically acceptable saltsof such acids and esters are active antibacterial agents.

Preferred acids, esters and salts of formula IV are those in which Z isC₁ -C₆ alkyl and acyl group R is selected from the acyl groups definedabove under (i) to (xvi). The acyl groups mentioned as being preferredgroups within categories (i) to (xvi) are also preferred in thecompounds defined by general formula IV.

More preferred acids, esters and salts of formula IV are those in whichZ is C₁ -C₆ alkyl and acyl group R is ##STR58## in which Ar' is aradical of the formula ##STR59## in which R^(m), R^(n) and R^(o) arealike or different and each is hydrogen, hydroxy, (lower)alkyl, cyano,(lower)alkoxy, chloro, bromo, iodo, fluoro, trifluoromethyl, nitro,amino, (lower)alkylamino, di(lower)alkylamino, (lower)alkylamino,(lower)alkanoyl, (lower)alkanoyloxy such as p-acetoxy or phenyl and Y isamino or a group obtained by reacting the amino group with acetaldehyde,formaldehyde or acetone; fluoro; chloro; bromo; iodo; hydroxy;(lower)alkanoyloxy; carboxy; guanidino; 3-guanyl-1-ureido;3-(2-furoyl)ureido; 3-benzoylureido; sulfo; sulfoamino; ureido;thioureido; (lower)alkoxy; cyano; cyanamino; or indanyloxycarbonyl.

Other preferred acids, esters and salts of formula IV are those in whichZ is C₁ -C₆ alkyl and R is ##STR60## wherein Ar' is phenyl,p-hydroxyphenyl, 4-hydroxy-3,5-dichlorophenyl, 3-chloro-4-hydroxyphenylo-, m- or p-aminomethylphenyl, 2-thienyl, 3-thienyl, 1-cyclohexenyl or1,4-cyclohexadienyl and Y is amino, carboxy or hydroxy.

Other preferred compounds of formula IV are those wherein Z is C₁ -C₆alkyl and R is an acyl group of the formula ##STR61##

Other preferred compounds of formula IV are those wherein Z is C₁ -C₆alkyl and R is an acyl group of the formula ##STR62## wherein U and Vare alike or different and each is hydrogen, chloro or fluoro; ##STR63##

A most preferred group of compounds are those acids defined by formulaIV wherein Z is C₁ -C₆ alkyl and R is α-carboxyphenylacetyl,cyanoacetyl, α-amino-α-(p-hydroxyphenyl)acetyl,α-amino-α-(3-chloro-4-hydroxyphenyl)acetyl,α-amino-α-(3,5-dichloro-4-hydroxyphenyl)acetyl,α-amino-α-(2-thienyl)acetyl, α-amino-α-(3-thienyl)acetyl,α-amino-α-(1-cyclohexenyl)acetyl,α-amino-α-(1,4-cyclohexadienyl)-acetyl, α-hydroxyacetal,α-hydroxy-α-(2-thienyl)acetyl, α-hydroxy-α-(3-thienyl)acetyl,α-hydroxy-α-(1-cyclohexenyl)acetyl,α-hydroxy-α-(1,4-cyclohexadienyl)acetyl, α-carboxy-α-(2-thienyl)acetyl,α-carboxy-α-(3-thienyl)-acetyl, α-carboxy-α-(1-cyclohexenyl)acetyl,α-carboxy-α-(1,4-cyclohexadienyl)acetyl,α-indanyloxycarbonyl-α-phenylacetyl 1-(1H)tetrazolyl,4-pyridylthioacetyl, 2-thienylacetyl, 3-thienylacetyl,1-cyclohexenylacetyl, 1,4-cyclohexadienylacetyl,o-aminomethylphenylacetyl, 1-aminocyclohexylcarbonyl,2,6-dimethoxybenzoyl, sydnoneacetyl or α-azidophenylacetyl, or apharmaceutically acceptable salt thereof.

Another most preferred group of compounds are the D-isomers of thoseacids defined by formula IV wherein Z is C₁ -C₆ alkyl and R isα-amino-α-(p-hydroxyphenyl)-acetyl,α-amino-α-(3-chloro-4-hydroxyphenyl)acetyl,α-amino-α-(3,5-dichloro-4-hydroxyphenylacetyl,α-amino-α-(2-thienyl)acetyl, α-amino-α-(3-thienyl)acetyl,α-amino-α-(1,-cyclohexenyl)acetyl,α-amino-α-(1,4-cyclohexadienyl)acetyl, α-hydroxyacetyl,α-hydroxy-α-(2-thienyl)-acetyl, α-hydroxy-α-(3-thienyl)acetyl,α-hydroxy-α-(1-cyclohexenyl)acetyl orα-hydroxy-α-(1,4-cyclohexadienyl)-acetyl, or a pharmaceuticallyacceptable salt thereof.

A most preferred group of compounds of formula IV are the acids in whichZ is C₁ -C₆ alkyl and R is phenoxyacetyl, or a pharmaceuticallyacceptable salt thereof.

Another most preferred group of compounds of formula IV are the acids inwhich Z is C₁ -C₆ alkyl and R is phenyl acetyl, or a pharmaceuticallyacceptable salt thereof.

A still further most preferred group of compounds of formula IV are theacids in which Z is C₁ -C₆ alkyl and R is α-aminophenylacetyl, or apharmaceutically acceptable salt thereof. The isomers of these compoundsin which the α-carbon atom of the 7-acyl group is of the D-series are ofparticular importance due to their combination of good antibacterialactivity and oral absorption.

The most preferred compounds of formula IV are those acids, esters andsalts where Z is methyl.

The present invention further provides various novel intermediatesuseful in the synthesis of the pharmacologically active O-2-isocephemantibacterial agents described above.

Preferred embodiments of the present invention are the novelintermediates having the formula ##STR64## wherein R" is hydrogen or aneasily cleavable ester carboxyl-protecting group, and salts thereof.

Other preferred intermediates are those having the formula ##STR65##wherein W is C₁ -C₁₀ alkyl, especially methyl, or aralkyl, especiallybenzyl or phenethyl, and R" is hydrogen or an easily cleavable estercarboxyl-protecting group, and salts thereof.

The most preferred intermediates of formula VI are those in which W ismethyl.

Other preferred intermediates are the compounds having the formula##STR66## wherein R" is hydrogen or an easily cleavable estercarboxyl-protecting group, and salts thereof.

Other preferred intermediates are the compounds having the formula##STR67## where W is C₁ -C₁₀ alkyl, especially methyl, or aralkyl,especially benzyl or phenethyl, and R" is hydrogen or an easilycleavable ester carboxyl-protecting group, and salts thereof. The mostpreferred intermediates of formula VIII are those in which W is methyl.

Still other preferred embodiments of the present invention are the novelintermediates having the formula ##STR68## wherein W is C₁ -C₁₀ alkyl,preferably methyl, or aralkyl, preferably benzyl or phenethyl, and R' isan easily cleavable ester carboxyl-protecting group, and salts thereof.The (lower)alkyl substituent is preferably methyl. Formulae IX and X arecondensed structural formulae and compounds IX and X as well as theother compounds of this invention may also be represented by structuralformulae of the type, e.g. ##STR69##

The intermediates of formulae V to X may be in the form of the freecarboxylic acid or a salt thereof or in the form where the carboxylgroup is protected in a conventional manner such as preferably byesterification. The protecting group is selected so that it may beremoved by methods which do not result in any appreciable destruction ofthe remaining portion of the molecule. Preferred carboxyl protectinggroups are the easily cleavable esters as defined above including inparticular benzhydryl, p-nitrobenzyl, trichloroethyl, silyl includingespecially trimethylsilyl, phenacyl, p-methoxybenzyl, acetonyl,(lower)-alkyl such as methyl, t-butyl or ethyl, benzyl, triphenylmethyl,phthalidyl, indanyl, methoxymethyl, acetoxymethyl and pivaloyloxymethyl.The most preferred esters of formulae V to X are the pivaloyloxymethyl,methoxymethyl, phthalidyl, indanyl and acetoxymethyl esters, and saltsthereof. The carboxyl-protecting group may be split off when desired bymethods known per se, e.g. by mild acid or base hydrolysis, catalytichydrogenation, irradiation with ultraviolet light, or reduction withchemical reducing agents. It will be appreciated that esterification isonly a preferred method for blocking the carboxyl group and that othercarboxyl-protected forms of the above intermediates, e.g. easilycleavable amides or anhydrides, are also intended to be included withinthe scope of the invention.

According to the present invention there is also provided a process forpreparing O-2-isocephem compounds of the formula ##STR70## wherein Rrepresents an acyl group, and easily cleavable esters andpharmaceutically acceptable salts thereof; which process comprises theconsecutive steps of

(1) reacting an ester of the formula ##STR71## wherein R' is an easilycleavable ester carboxyl-protecting group with cinnamaldehyde in aninert organic solvent in the presence of a drying agent or withazeotropic removal of water to produce an imine of the formula ##STR72##wherein R' is as defined above;

(2) reacting said imine of formula XII with an azidoacetyl halide or anazidoacetic mixed anhydride in the presence of an organic base toproduce a cis-β-lactam compound of the formula ##STR73## wherein R' isas defined above;

(3) subjecting said β-lactam compound of formula IV to ozonolysis toproduce an aldehyde of the formula ##STR74## wherein R' is as definedabove;

(4) selectively reducing said aldehyde of formula XXVIII to thecorresponding alcohol of the formula ##STR75## wherein R' is as definedabove;

(5) reacting the alcohol of formula XIII with an acid anhydride,preferably acetic anhydride or trifluoroacetic anhydride, in thepresence of a Lewis acid to produce a compound of the formula ##STR76##wherein R' is as defined above and An represents the residue of the acidanhydride;

(6) subjecting the compound of formula XIV to acid hydrolysis to producea compound of the formula ##STR77## wherein R' is as defined above;

(7) reacting the compound of formula XV with an amine, preferably asaturated cyclic secondary amine, to produce an enamine compound of theformula ##STR78## wherein R' is as defined above and X" is the residueof the amine;

(8) reacting the enamine of formula XVI with a (lower)alkylsulfonatingagent in the presence of an acid acceptor to produce a derivative of theformula ##STR79## wherein R' and X" are as defined above;

(9) subjecting said derivative of formula XVII to acid hydrolysis toproduce an enol intermediate of the formula ##STR80## wherein R' is asdefined above;

(10) cyclizing said enol intermediate by treatment with base to producean azido O-2-isocephem intermediate of the formula ##STR81## wherein R'is as defined above;

(11) selectively reducing said azido intermediate of formula Va toproduce a 7-amino O-2-isocephem intermediate of the formula ##STR82##wherein R' is as defined above; and

(12) N-acylating said 7-amino intermediate of formula VIIa or a saltthereof with an acylating acid of the formula

    R--COOH

wherein R is as defined above, or with its functional equivalent as anacylating agent for a primary amine, to produce a 7-acylamidoO-2-isocephem-4-carboxylic acid derivative of the formula ##STR83##wherein R and R' are as defined above, or a pharmaceutically acceptablesalt thereof and, if desired, performing one or more of the additionalsteps of

(13) removing by methods known per se the carboxyl-protecting group R'to produce the corresponding free acid compound of formula II;

(14) converting the free acid compound of formula II to aphysiologically hydrolyzed ester thereof by methods known per se; or

(15) converting the free acid compound of formula II or aphysiologically hydrolyzed ester thereof to a pharmaceuticallyacceptable salt thereof by methods known per se.

A variation of the above-described process for preparing theO-2-isocephem antibacterial agents of formula II and easily cleavableesters and pharmaceutically acceptable salts thereof involves reactingthe alcohol of formula XIII with a (lower)alkylsulfonating agent in thepresence of an acid acceptor to produce a derivative of the formula##STR84## wherein R' is an easily cleavable ester carboxyl-protectinggroup. Compound XVIII is then reacted with an acid anhydride in thepresence of a Lewis acid to produce a compound of the formula ##STR85##wherein R' is as defined above. Compound XIX is then subjected to basehydrolysis to form an enolate of the formula ##STR86## wherein R' is asdefined above. Enolate XX then cyclizes on removal of water and exposureto a dipolar aprotic solvent such as dimethylsulfoxide to form the7-azido O-2-isocephem intermediate of formula Va which is subsequentlyreduced, N-acylated and optionally de-blocked according to the processdescribed above to produce the desired compound of formula II or aneasily cleavable ester or pharmaceutically acceptable salt thereof.

A still further alternative process for preparing compounds of formulaII involves reacting the compound of the formula ##STR87## wherein R' isan easily cleavable ester carboxyl-protecting group with a(lower)alkylsulfonating agent in the presence of an acid acceptor toproduce a derivative of the formula ##STR88## wherein R' is as definedabove. Compound XIX is then treated with base to produce an enolate offormula XX which cyclizes to the 7-azido O-2-isocephem intermediate Vawhich is reacted as described above to produce the desired compound offormula II or an easily cleavable ester or pharmaceutically acceptablesalt thereof.

A preferred embodiment of the present invention is the process ofpreparing a 7-acylamido O-2-isocephem-4-carboxylic acid compound havingthe formula ##STR89## wherein R represents an acyl group, and easilycleavable esters and pharmaceutically acceptable salts thereof; whichprocess comprises N-acylating a 7-amino O-2-isocephem intermediate ofthe formula ##STR90## wherein R" is hydrogen or an easily cleavableester carboxyl-protecting group R', or a salt thereof, with an acylatingacid of the formula

    R--COOH

wherein R is as defined above, or with its functional equivalent as anacylating agent for a primary amine, to produce a 7-acylamidoO-2-isocephem-4-carboxylic acid derivative of the formula ##STR91##wherein R and R" are as defined above, or a pharmaceutically acceptablesalt thereof and, if desired, performing one or more of the additionalsteps of

(1) removing by methods known per se the carboxyl-protecting group R' toproduce the corresponding free acid compound of formula II;

(2) converting the free acid compound of formula II to a physiologicallyhydrolyzed ester thereof by methods known per se; or

(3) converting the free acid compound of formula II or a physiologicallyhydrolyzed ester thereof to a pharmaceutically acceptable salt thereofby methods known per se.

Another preferred embodiment of the present invention is the process ofpreparing a 7-amino O-2-isocephem intermediate of the formula ##STR92##wherein R" is hydrogen or an easily cleavable ester carboxyl-protectinggroup, or a salt thereof; which process comprises selectively reducing a7-azido intermediate of the formula ##STR93## wherein R' is an easilycleavable ester carboxyl-protecting group and, if desired, removingprotecting group R' by methods known per se to produce the correspondingfree acid intermediate of formula VII and, if desired, converting thefree acid form of intermediate VII to a salt thereof by methods knownper se.

Another preferred embodiment of the present invention is the process ofpreparing a 7-azido O-2-isocephem intermediate of the formula ##STR94##wherein R" is hydrogen or an easily cleavable ester carboxyl-protectinggroup, or a salt thereof; which process comprises cyclizing by treatmentwith base an anol intermediate of the formula ##STR95## wherein R' is aneasily cleavable ester carboxyl-protecting group and, if desired,removing protecting group R' by methods known per se to produce thecorresponding free acid intermediate of formula V and, if desired,converting the free acid to a salt thereof by methods known per se.

Still another preferred embodiment of the present invention is theprocess of preparing a 7-azido O-2-isocephem intermediate of the formulaV wherein R" is hydrogen or an easily cleavable estercarboxyl-protecting group, or a salt thereof; which process comprisessubjecting a compound of the formula ##STR96## wherein R' is an easilycleavable ester carboxyl-protecting group to base hydrolysis andcyclization to produce the desired carboxyl-protected intermediate offormula V and, if desired, removing protecting group R' by methods knownper se to produce the corresponding free acid intermediate of formula Vand, if desired, converting the free acid to a salt thereof by methodsknown per se.

A further preferred embodiment of the present invention is a process forthe preparation of an enol intermediate of the formula ##STR97## whereinR' is an easily cleavable ester carboxyl-protecting group; which processcomprises subjecting to acid hydrolysis a derivative of the formula##STR98## wherein R' is as defined above and X" is the residue of anamine.

The ester starting material of formula XI may be prepared by the generalmethod as illustrated in more detail in the disclosure and examplesbelow for the case in which R' is ethyl. The ethyl ester of formula XImay be prepared by the following reaction sequence: ##STR99## Byreplacing the ethyl glycinate in the above process with other desiredeasily cleavable esters, starting materials having other suitablecarboxyl-protecting groups may be prepared.

Conversion of ester XI to the Schiff base XII may be effected bycondensing the ester with cinnamaldehyde in an inert organic solvent.The Schiff base usually forms quite readily at room temperature butcatalysis with the HCl salt of the amine may be used, if desired, toaccelerate the reaction. The condensation reaction is carried out in asuitable inert organic solvent, e.g. benzene, ether or methylenechloride, in the presence of a drying agent, e.g. Na₂ SO₄, MgSO₄,molecular sieves, etc., or, alternatively, by removing waterazeotropically as with benzene.

The imine of formula XII is then condensed with an azidoacetyl halide,preferably azidoacetyl chloride, or an azidoacetyl mixed anhydride inthe presence of an organic base, preferably a tertiary amine such as atrialkylamine or pyridine. The reaction may be conducted in an inertorganic solvent which may advantageously be a hydrocarbon or halogenatedsolvent. A most preferred solvent for this step is methylene chloride.Best results are obtained when the reaction mixture is cooled to about0°-5° C. At the conclusion of the reaction, the Schiff base solution isdried as with Na₂ SO₄ and preferably evaporated to dryness. Compound IVis obtained as a mixture of diastereoisomers having the formulae##STR100##

The stereochemistry of the azido and stryrryl substituents of compoundIV has been shown by NMR to be exclusively cis. The diastereoisomers ofIV need not be separated for use in the next step of the reactionsequence.

After formation of the cis β-lactam compound IV, the ester protectinggroup R' may, if desired, be removed, e.g. by saponification, and thecorresponding free acid esterified to incorporate into compound IVanother ester moiety R'. This optional transesterification step mayconveniently be used to convert an ester such as a (lower)alkyl ester ofIV to a more labile and easily removable ester such as a benzyl ester.Preferably, however, the desired easily removable ester group isincorporated into starting material XI so as to avoid the necessity of alater transesterification step.

The cis β-lactam ester of formula IV is subjected to ozonolysis in thenext step of the process to produce the aldehyde of formula XVIII.Ozonolysis is conveniently conducted in an inert organic solvent, e.g.methylene chloride, with cooling, e.g. -50° to -80° C. Followingozonolysis a reagent such as dimethylsulfide, dimethylsulfoxide ortriethylamine is preferably added to the reaction mixture to decomposethe ozonide formed initially in the reaction.

Aldehyde XVIII is next selectively reduced to form the alcohol compoundXIII. Convenient reducing agents for this step include diborane or metalhydrides such as sodium borohydride, lithium borohydride or zincborohydride. The reduction is conducted in an inert organic solventwhich most advantageously is a (lower)alkanol such as ethanol ormethanol or tetrahydrofuran and preably with cooling to a temperature ofabout 0° to -10° C.

According to one reaction route, alcohol XIII is next converted tocompound XIV by treatment with an acid anhydride, preferably aceticanhydride or trifluoroacetic anhydride, in the presence of a Lewis acid,preferably an acid selected from zinc chloride, trityl fluoroborate,triethyl oxonium fluoroborate, titanium tetrachloride, boron trifluorideor stannous chloride. The three reactants in this step are preferablyused in equimolar amounts and the reaction may conveniently be performedat room temperature.

Compound XIV is hydrolyzed to alcohol XV by treatment with acid,conveniently an aqueous solution of a mineral acid, e.g. hydrochloricacid. The hydrolysis is advantageously conducted at reflux temperatures.

In the next step of this reaction route, alcohol XV is reacted with anamine, preferably a saturated cyclic secondary amine such as morpholine,piperidine or pyrrolidine, to produce the enamine compound of formulaXVI. This step is conveniently effected by refluxing the alcohol andamine in an inert organic solvent, e.g. benzene, in the presence of anorganic acid, e.g. acetic acid.

Enamine XVI is then reacted with a (lower)alkyl-sulfonating agent,preferably a methanesulfonating agent such as methanesulfonic anhydrideor methanesulfonyl chloride, in the presence of an acid acceptor,preferably a tertiary amine such as a trialkylamine, e.g. triethylamine,or pyridine to produce a derivative of formula XVII. The reaction may beconducted in an inert organic solvent, e.g. methylene chloride, and isconveniently carried out at room temperature.

The derivative of formula XVII is next hydrolyzed by treatment with acidto the enol intermediate of formula IX. Advantageously the hydrolysis iscarried out by refluxing an aqueous solution of a mineral acid such ashydrochloric acid with compound XVII.

Enol intermediate IX is cyclized to the azido O-2-isocephem intermediateVa by treatment with a base. Examples of suitable bases for this stepinclude an alkali metal hydride, most preferably NaH, in a suitableorganic solvent, e.g. dimethylsulfoxide or dimethylformamide,triethylamine in an inert organic solvent such as methylene chloride,chloroform or (lower)alkanols, e.g. methanol or ethanol and sodium orpotassium acetate in a suitble solvent such as dimethylformamide.Generally it is preferred to employ the azido O-2-isocephem in itscarboxyl-protected form in preparing the active O-2-isocephemantibacterial agents of formula II. If desired, however, the ester groupof intermediate Va may be removed by methods known per se to produce thefree acid form of compound Va. The ester or free acid forms of theazido-isocephem intermediate may also be optionally converted by knownmethods to salts thereof.

In an alternative reaction route, alcohol XIII is reacted with a(lower)alkylsulfonating agent, preferably a methanesulfonating agentsuch as methanesulfonic anhydride or methanesulfonyl chloride, in thepresence of an acid acceptor, preferably a tertiary amine such as atrialkylamine, e.g. triethylamine, or pyridine, to form the derivativeXVIII. The reaction may be conducted in an inert organic solvent, e.g.methylene chloride, and is conveniently carries out at room temperature.Compound XVIII is then converted by treatment with an acid anhydride,preferably acetic anhydride or trifluoroacetic anhydride, in thepresence of a Lewis acid, preferably an acid selected from zincchloride, trityl fluoroborate, triethyl oxonium fluoroborate, titaniumtetrachloride, boron trifluoride or stannous chloride, to compound XIXwhich may then be subjected to base hydrolysis to form the enolateintermediate XX. While the nature of the base is not critical in thehydrolysis step, advantageously the base is an alkali metal hydroxidesuch as sodium or potassium hydroxide. Enolate intermediate XX thencyclizes on removal of water and exposure to a dipolar aprotic solventsuch as dimethylsulfoxide to form the 7-azido O-2-isocephem intermediateVa.

In still another alternative reaction route, alcohol XV is converted toa (lower)alkylsulfonyl derivative XIX by reaction with a(lower)alkylsulfonating agent, preferably a methanesulfonating agentsuch as methanesulfonic anhydride or methanesulfonyl chloride, in thepresence of an acid acceptor, preferably a tertiary amine base such as atrialkylamine, e.g. triethylamine, or pyridine, in an inert organicsolvent, e.g. methylene chloride, and preferably at about roomtemperature. Compound XIX is subjected to base hydrolysis to formenolate XX which may be cyclized as described above to form the 7-azidoO-2-isocephem intermediate Va.

Azido-isocephem intermediate Va prepared according to any of the aboveprocedures is selectively reduced to the 7-amino O-2-isocephemintermediate VIIa. Preferred reducing agents for this step includechemical reducing agents selected from zinc and ammonium chloride in aninert organic solvent, e.g. methanol, and hydrogen sulfide andtriethylamine in an inert organic solvent, e.g. methylene chloride.Catalytic hydrogenation may also be employed with such selectivehydrogenation catalysts as noble metals (most preferably palladium orplatinum), noble metal oxides (most preferably palladium oxide orplatinum oxide), or aluminum amalgam or Raney nickel, said catalystsbeing optionally supported on a conventional carrier such as carbon,diatomaceous earth, etc. Preferred solvents for catalytic hydrogenationare non-reducible inert solvents such as methanol, ethanol and ethylacetate. Hydrogenation is preferably conducted at room temperature andat atmospheric or slightly elevated pressure. As in the case of the7-azido O-2-isocephem intermediate discussed above, compound VIIa may,if desired, be converted by methods known per se to the correspondingfree carboxylic acid form or to a salt of either the esterified or freeacid form. As an alternative to the stepwise reduction of the azidogroup and removal of the ester carboxyl-protecting group, it is alsopossible by choice of proper reduction conditions and protecting groupsto simultaneously reduce the azido group and ester protecting group.Thus if a mild hydrogenation catalyst is used such as 10% Pd-on-charcoalor a mild chemical reducing agent such as H₂ S and triethylamine, theazido group is reduced to an amine but a benzyl protecting group isuntouched. If a more active catalyst, however, such as 30%Pd-on-diatomaceous earth is employed, both azido and benzyl groups arereduced.

Compound VIIa or an easily cleavable ester or salt thereof is of useprimarily as an intermediate in preparing the active N-acyl derivativesof formula II. Compound VIIa upon conversion to the free carboxylic acid(or a physiologically hydrolyzed ester or pharmaceutically acceptablesalt of said acid or ester) also possesses antibacterial activity per seagainst Gram-positive and Gram-negative bacteria.

The 7-acylamido O-2-isocephem compounds of general formula II may beprepared by N-acylation according to methods known per se of the 7-aminogroup of intermediate VIIa or VII. The preferred methods for preparingthe active end-products of formula II will be discussed in more detailbelow.

According to another aspect of the present invention, there is provideda process for preparing O-2-isocephem compounds of the formula##STR101## wherein R represents an acyl group and W is C₁ -C₁₀ alkyl,preferably methyl, or aralkyl, preferably benzyl or phenethyl, andeasily cleavable esters and pharmaceutically acceptable salts thereof;which process comprises the consecutive steps of

(1) reacting a ketal amine of the formula ##STR102## wherein R'represents an easily cleavable ester carboxyl-protecting group and W isC₁ -C₁₀ alkyl or aralkyl with cinnamaldehyde in an inert organic solventin the presence of a drying agent or with azeotropic removal of water toproduce an imine of the formula ##STR103## wherein R' and W are asdefined above;

(2) reacting said imine with an azidoacetyl halide or an azidoaceticmixed anhydride in the presence of a base to produce a cis-β-lactamcompound of the formula ##STR104## wherein R' and W are as definedabove;

(3) subjecting said β-lactam compound of formula XXIV to ozonolysis toproduce an aldehyde of the formula ##STR105## wherein R' and W are asdefined above;

(4) selectively reducing the aldehyde of formula XXV to thecorresponding alcohol of the formula ##STR106## wherein R' and W are asdefined above;

(5) reacting said alcohol with a (lower)alkylsulfonating agent in thepresence of an acid acceptor to produce a derivative of the formula##STR107## wherein R' and W are as defined above;

(6) subjecting said derivative of formula XXVII to acid hydrolysis toform an enol of the formula ##STR108## wherein R' and W are as definedabove;

(7) cyclizing the enol of formula X by treatment with base to produce anazido O-2-isocephem intermediate of the formula ##STR109## wherein R'and W are as defined above: (8) selectively reducing the azidoO-2-isocephem intermediate of formula VIa to produce a 7-aminoO-2-isocephem intermediate of the formula ##STR110## wherein R' and Ware as defined above; and

(9) N-acylating said 7-amino intermediate of formula VIIIa or a saltthereof with an acylating acid of the formula

    R--COOH

wherein R represents an acyl group, or with its functional equivalent asan acylating agent for a primary amine, to produce a 7-acylamidoO-2-isocephem-4-carboxylic acid derivative of the formula ##STR111##wherein R, W and R' are as defined above, or a pharmaceuticallyacceptable salt thereof and, if desired, performing one or more of theadditional steps of

(10) removing by methods known per se the carboxyl-protecting group R'to produce the corresponding free acid compound of formula III;

(11) converting the free acid compound of formula III to aphysiologically hydrolyzed ester thereof by methods known per se; or

(12) converting the free acid compound of formula III or aphysiologically hydrolyzed ester thereof to a pharmaceuticallyacceptable salt thereof by methods known per se.

A preferred embodiment of the present invention is the process ofpreparing a 7-acylamido O-2-isocephem compound having the formula##STR112## wherein R represents an acyl group and W is C₁ -C₁₀ alkyl,preferably methyl, or aralkyl, preferably benzyl or phenethyl, andeasily cleavable esters and pharmaceutically acceptable salts thereof;which process comprises N-acylating a 7-amino O-2-isocephem intermediateof the formula ##STR113## wherein R" is hydrogen or an easily cleavableester carboxyl-protecting group R' and W is as defined above, or a saltthereof, with an acylating acid of the formula

    R--COOH

wherein R is as defined above, or with its functional equivalent as anacylating agent for a primary amine, to produce a 7-acylamidoO-2-isocephem-4-carboxylic acid derivative of the formula ##STR114##wherein R, R" and W are as defined above, or a pharmaceuticallyacceptable salt thereof and, if desired, performing one or more of theadditional steps of

(1) removing by methods known per se the carboxyl-protecting group R' toproduce the corresponding free acid compound of formula III; (2)converting the free acid compound of formula III to a physiologicallyhydrolyzed ester thereof by methods known per se; or

(3) converting the free acid compound of formula III or aphysiologically hydrolyzed ester thereof to a pharmaceuticallyacceptable salt thereof by methods known per se.

Another preferred embodiment of the present invention is the process ofpreparing a 7-amino O-2-isocephem intermediate of the formula ##STR115##wherein W is C₁ -C₁₀ alkyl, preferably methyl, or aralkyl, preferablybenzyl or phenethyl, and R" is hydrogen or an easily cleavable estercarboxyl-protecting group, or a salt thereof; which process comprisesselectively reducing a 7-azido intermediate of the formula ##STR116##wherein R' is an easily cleavable ester carboxyl-protecting group and Wis as defined above, or a salt thereof and, if desired, removingprotecting group R' by methods known per se to produce the correspondingfree acid intermediate of formula VIII and, if desired, converting thefree acid to a salt thereof by methods known per se.

Another preferred embodiment of the present invention is the process ofpreparing a 7-azido O-2-isocephem intermediate of the formula ##STR117##wherein W is C₁ -C₁₀ alkyl, preferably methyl, or aralkyl, preferablybenzyl or phenethyl, and R" is hydrogen or an easily cleavable estercarboxyl-protecting group, or a salt thereof; which process comprisescyclizing by treatment with base an enol intermediate of the formula##STR118## wherein W is as defined above and R' is an easily cleavableester carboxyl-protecting group and, if desired, removing protectinggroup R' by methods known per se to produce the corresponding free acidintermediate of formula VI and, if desired, converting the free acidintermediate to a salt thereof by methods known per se.

A further preferred embodiment of the present invention is a process forthe preparation of an enol intermediate of the formula ##STR119##wherein W is C₁ -C₁₀ alkyl, preferably methyl, or aralkyl, preferablybenzyl or phenethyl, and R' is an easily cleavable estercarboxyl-protecting group; which process comprises subjecting to acidhydrolysis a derivative of the formula ##STR120## wherein R' and W areas defined above

The ketal amine starting material of formula XXII may be prepared by thefollowing reaction sequence: ##STR121##

The ester oxime may be prepared according to the general methoddescribed by H. Adkins and J. Reeve, J.A.C.S., 60, 1328 (1939). Thecarbonyl group of the ester oxime is protected by ketalization withethylene glycol in the presence of acid, e.g. p-toluenesulfonic acid.The ketal oxime is then selectively reduced as with aluminum amalgam bythe procedure described by D. J. Drinkwater and P. W. G. Smith in J.Chem. Soc. (C), 1305, (1971). The product may be recovered as an acidaddition salt, e.g. HCl salt, which may be basified to give ketal aminestarting material XXII.

An alternative procedure for preparing ketal amine starting materials offormula XXII having less labile ester groups, e.g. (lower)alkyl esterssuch as ethyl or methyl, comprises selectively reducing the oxime groupof an ester oxime of the formula ##STR122## and subsequently protectingthe carbonyl group by ketalization. The reduction step may beconveniently accomplished by catalytic hydrogenation and theketalization step is carried out as described above and in the exampleswhich follow.

Conversion of the ketal amine XXII to alcohol XXVI is effected asdescribed above in connection with the process for preparing compoundsof formula II. Compound XXIV is obtained as a mixture ofdiastereoisomers having the formulae ##STR123##

The stereochemistry of the azido and styrryl substituents of compoundXXIV has been shown by NMR to be exclusively cis. The diastereoisomersof XXIV need not be separated for use in the ozonolysis step of thereaction sequence.

Alcohol XXVI is reacted with a (lower)alkylsulfonating agent, preferablya methanesulfonating agent such as methanesulfonic anhydride ormethansulfonyl chloride, in the presence of an acid acceptor, preferablya tertiary amine, e.g. triethylamine, or pyridine, to produce thederivative of formula XXVII. The reaction may be conducted in an inertorganic solvent, e.g. methylene chloride, and is conveniently carriedout at room temperature.

Enol intermediate X is produced by hydrolyzing compound XXVII underacidic conditions. Examples of suitable reagents for the ketalhydrolysis step are trifluoroacetic acid, a solution of BF₃ etherate and50% perchloric acid in acetone.

Following the hydrolysis step, the enol intermediate X is cyclized bytreatment with base to produce the azido O-2-isocephem intermediate VIa.Examples of suitable bases for the cyclization step include an alkalimetal hydride, e.g. NaH, in a suitable organic solvent, e.g.dimethylsulfoxide or dimethylformamide, triethylamine in an inertorganic solvent such as methylene chloride, chloroform or a(lower)alkanol, e.g. methanol or ethanol, and sodium or potassiumacetate in a suitable solvent such as dimethylformamide. Generally it ispreferred to employ the azido O-2-isocephem in the carboxyl-protectedform in preparing the active O-2-isocephem antibacterial agents offormula III. If desired, however, the ester protecting group ofintermediate VIa may be cleaved by methods known per se to produce thefree carboxylic acid form of compound VIa. The ester or free acid formsof the azido isocephem intermediate may also be optionally converted byknown methods to salts thereof.

Azido-isocephem intermediate VIa is next selectively reduced to the7-amino O-2-isocephem intermediate VIIIa by catalytic hydrogenation withsuch selective hydrogenation catalysts as Raney nickel, aluminumamalgam, noble metals such as preferably platinum or palladium(optionally on a carrier such as carbon), or noble metal oxides such aspreferably platinum oxide or palladium oxide or by the use of chemicalreducing agents such as most preferably zinc and ammonium chloride in aninert organic solvent, e.g. methanol, or hydrogen sulfide andtriethylamine in an inert organic solvent, e.g. methylene chloride.Preferred solvents for catalytic hydrogenation are such non-reducibleinert solvents as methanol, ethanol and ethyl acetate. Hydrogenation ispreferably conducted at room temperature and at atmospheric or slightlyelevated pressure. As in the case of the 7-azido O-2-isocephemintermediate VIa discussed above, compound VIIIa may, if desired, beconverted by methods known per se to the corresponding free carboxylicacid form or to a salt of either the esterified or free acid form.

The compounds of formula VIIIa or easily cleavable esters or saltsthereof are of use primarily as intermediates in preparing the activeN-acyl derivatives of formula III. Compounds of formula VIIIa, however,upon conversion to the free carboxylic acid (or a physiologicallyhydrolyzed ester or pharmaceutically acceptable salt of said acid orester) also possess antibacterial activity per se against a variety ofGram-positive and Gram-negative bacteria.

The 7-acylamido O-2-isocephem compounds of formulae II and III areprepared by N-acylation according to known methods of the 7-amino groupof intermediate VIIa or VIIIa with an acylating acid of the formula

    R--COOH

wherein R is an acyl group, or with its functional equivalent as anacylating agent for a primary amino group. The acylating agents forpreparing the products of formulae II and III are known, readilypreparable by known methods or described herein.

Intermediates VIIa and VIIIa may be acylated either in the form of thefree carboxylic acid (or salt thereof) or as an easily cleavable ester(or acid addition salt thereof). The procedures for preparing esters ofcarboxylic acids are disclosed in the literature and are well-known tothose skilled in the art of penicillin and cephalosporin chemistry.Methods for preparing certain of the more preferred easily cleavableesters, i.e. the pivaloyloxymethyl, acetoxymethyl, methoxymethyl,acetonyl and phenacyl esters, are disclosed in U.S. Pat. No. 3,284,451and in U.K. Pat. No. 1,229,453. Preparation of the phthalidyl ester isdescribed in South African Application 72/3799. The free acid form ofintermediate VIIa and VIIIa may also be converted to a silyl ester, e.g.trimethylsilyl ester, as by the methods described in the literature,e.g. U.S. Pat. No. 3,249,622. The silyl ester carboxyl-protecting groupmay be easily removed following the acylation reaction by hydrolysis oralcoholysis.

Prior to the acylation reaction, any reactive substituents on theacylating acid or derivative thereof, e.g. hydroxy, carboxyl ormercapto, may be protected by use of suitable protecting or blockinggroups which are well-known to those skilled in the art of β-lactamchemistry, e.g. as by acylation or silylation. When the acylating agentcontains an amino functional group in the acyl moiety, the amino groupis protected by a conventional amino-blocking group which may be readilyremoved at the conclusion of the reaction. Examples of suitableamino-protecting or blocking groups include t-butoxycarbonyl,carbobenzyloxy, 2-hydroxy-1-naphthcarbonyl, trichloroethoxycarbonyl,2-ethoxycarbonyl-1-methylvinyl and 2-methoxycarbonyl-1-methylvinyl. Aparticularly valuable blocking group is a proton, as in the acylatingagent of the formula ##STR124## Preferred amino-protecting groups aret-butoxycarbonyl, carbobenzyloxy, the proton and a β-diketone orβ-ketoester as in U.K. Pat. No. 1,123,333 or U.S. Pat. Nos. 3,325,479and 3,316,247, e.g. methyl acetoacetate, or a β-ketoamide as in JapanPat. No. 71/24714. When the t-butoxycarbonyl, carbobenzyloxy,β-ketoester, β-diketone or β-ketoamide protecting groups are employed,it is preferred to convert the acylating acid containing the blockedamino group to a mixed anhydride, e.g. with ethyl or isobutylchloroformate, before reaction with compounds VIIa or VIIIa or an esteror salt thereof. After the acylation coupling reaction, theamino-protecting group and any other protecting group used may beremoved by methods known per se to form the desired product of formulaeII or III. Thus, for example, with respect to amino-protecting groups,the t-butoxycarbonyl group may be removed by use of formic acid, thecarbobenzyloxy group by catalytic hydrogenation, the2-hydroxy-1-naphthcarbonyl group by acid hydrolysis, thetrichloroethoxycarbonyl group by treatment with zinc dust in glacialacetic acid, the proton by neutralization, etc.

Acylation of a free amino group of a cephalosporin or penicillin nucleusis a well-known reaction, and any of the functional equivalents of thecarboxylic acid RCOOH commonly used in penicillin or cephalosporinchemistry as acylating agents for primary amino groups may be employedin acylating intermediate VIIa or VIIIa. Examples of suitable acylatingderivatives of the free acid include the corresponding acid anhydrides,mixed anhydrides, e.g. alkoxyformic anhydrides, acid halides, acidazides, active esters and active thioesters. The free acid may becoupled with compound VIIa or VIIIa after first reacting said free acidwith N,N'-dimethylchloroformininium chloride [cf. Great Britain Pat. No.1,008,170 and Novak and Weichet, Experientia XXI, 6, 360 (1965) ] or bythe use of enzymes or of an N,N'-carbonyldiimidazole or anN,N'-carbonylditriazole [cf. South African Specification 63/2684] or acarbodiimide reagent [especially N,N'-dicyclohexylcarbodiimide,N,N'-diisopropylcarbodiimide orN-cyclohexyl-N'-(2-morpholinoethyl)carbodiimide: cf. Sheehan and Hess,J.A.C.S., 77, 1967 (1955)], or of alkylylamine reagent [cf. R. Buijleand H. G. Viehe, Angew, Chem. International Edition, 3, 582, (1964)] orof an isoxasolium salt reagent [cf. R. B. Woodward, R. A. Olofson and H.Mayer, J. Amer. Chem. Soc., 83, 1010 (1961)], or of a ketenimine reagent[cf. C. L. Stevens and M. E. Munk, J. Amer. Chem. Soc., 80, 4065 (1958)]or of hexachlorocyclotriphosphatriazine orhexabromocyclotriphosphatriazine (U.S. Pat. No. 3,651,050) or ofdiphenylphosphoryl azide [DPPA; J. Amer. Chem. Soc., 94, 6203-6205(1972)] or of diethylphosphoryl cyanide [DEPC; Tetrahedron Letters No.18, pp. 1595-1598 (1973)] or of diphenyl phosphite [Tetrahedron LettersNo. 49, pp. 5047-5050 (1972)]. Another equivalent of the acid chlorideis a corresponding azolide, i.e., an amide of the corresponding acidwhose amide nitrogen is a member of a quasiaromatic five membered ringcontaining at least two nitrogen atoms, i.e., imidazole, pyrazole, thetriazoles, benzimidazole, benzotriazole and their substitutedderivatives. As an example of the general method for the preparation ofan azolide, N,N'-carbonyldiimidazole is reacted with a carboxylic acidin equimolar proportions at room temperature in tetrahydrofuran,chloroform, dimethylformamide or a similar inert solvent to form thecarboxylic acid imidazolide in practically quantitative yield withliberation of carbon dioxide and one mole of imidazole. Dicarboxylicacids yield dimidazolide. The by-product, imidazole, precipitates andmay be separated and the imidazolide isolated, but this is notessential. A preferred acylating agent for preparing 7-acylamidocompounds containing an α-amino substituent, e.g. α-aminophenyl,α-amino-α-thienyl, etc. is the N-carboxy anhydride (Leuch's anhydride).In this structure the group which activates the carboxyl group alsoserves to protect the amino group. Another preferred acylating agent forintroducing a side chain containing an α-amino functional group is theacid chloride hydrochloride, of the formula ##STR125## which also servesa dual function of carboxyl activation and amino protection. Mention wasmade above of the use of enzymes to couple the free acid with compoundVIIa and VIIIa. Included in the scope of such processes are the use ofan ester, e.g. the methyl ester, of that free acid with enzymes providedby various microorganisms, e.g. those described by T. Takahashi et al.,J.A.C.S., 94(11), 4035-4037 (1972) and by T. Nara et al., J. Antibiotics(Japan) 24(5), 321-323 (1971) and in U.S. Pat. No. 3,682,777. Aparticularly preferred coupling agent for coupling the acylating acidwith compound VIIa or VIIIa (or a salt or ester thereof) isN-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) as described inJ.A.C.S., 90, 823-824 and 1652-1653 (1968) and U.S. Pat. No. 3,455,929.

The particular process conditions, e.g. temperature, solvent, reactiontime, etc. selected for the coupling reaction are determined by thenature of the reactants and acylation method used and are known to thoseskilled in the art.

The acylating agents which may be used to form the N-acyl activecompounds of formulae II and III are known in the literature along withmethods for their synthesis. In those cases where the acylating agentcontains one or more asymmetric carbon atoms and thus exists inoptically active forms, the compounds obtained using such as acylatingagent are ordinarily obtained in racemic form. When the separate opticalisomers are desired, the acylating agent can be resolved in aconventional manner such as by reacting the free acid with cinchonine,strychnine, brucine or the like, fractionally crystallizing to separatethe diastereoisomeric salts and separately acidifying the solid phaseand the liquid phase to liberate the optical isomers.

The compounds of the present invention may be isolated in any of theways customarily employed for the isolation of correspondingcephalosporin compounds. Formation of the desired pharmaceuticallyacceptable carboxylic acid or acid addition salt is carried out by knownmethods, e.g. reaction of the acid (or ester in the case of acidaddition salts) with an appropriate base or acid.

At the conclusion of the acylation reaction, the product obtained may beconverted (before or after removal of any protecting groups) by methodsknown per se to another desired product of formulae II or III. Thus, thecompound of formula II or formula III in the form of the free acid or asalt thereof may be converted to known methods to the correspondingphysiologically hydrolyzed ester or pharmaceutically acceptable saltthereof. Similarly, the product of formula II or formula III in the formof an easily cleavable ester or salt thereof may be converted to thefree acid product or pharmaceutically acceptable salt thereof by removalof the esterifying group, e.g. by aqueous or enzymatic hydrolysis (aswith human or animal serum) or acidic or alkaline hydrolysis or bycatalytic hydrogenation or by treatment with sodium thiophenoxide astaught in U.S. Pat. No. 3,284,451.

The easily cleavable esters of the compounds of formulae II and III areuseful as intermediates in the production of the free acid product. Thepivaloyloxymethyl, acetoxymethyl, phthalidyl, indanyl and methoxymethylesters are also useful as active antibacterial agents since on oraladministration they are rapidly hydrolyzed to the active metabolite.These esters are of particular interest because they provide an oraladministration different rates and amounts of absorption and givediffering concentrations of the active antibacterial agent in blood andtissues.

In still another aspect of the present invention, there is provided aprocess for the preparation of an O-2-isocephem-3-carboxymethylenederivative of the formula ##STR126## wherein R represents an acyl groupand Z is hydrogen or the residue of an ester group, preferably C₁ -C₆alkyl, and easily cleavable esters and pharmaceutically acceptable saltsthereof; which process comprises

(1) reacting an O-2-isocephem compound of the formula ##STR127## whereinR represents an acyl group and R' is an easily cleavable estercarboxyl-protecting group with carbon dioxide in the present of a basein an inert organic solvent at a temperature in the range of about 0° to-80° C. to produce upon acidification the compound of the formula##STR128## wherein R and R' are as defined above, or a pharmaceuticallyacceptable salt thereof and optionally performing one or more of theadditional steps of

(a) removing by methods known per se the protecting group R' to producethe corresponding 4-carboxylic acid compound of formula XXII;

(b) converting the 4-carboxylic acid compound of formula XXII to aphysiologically hydrolyzed ester thereof by methods known per se; or

(c) converting by methods known per se the 4-carboxylic acid compound offormula XXII or a physiologically hydrolyzed ester thereof to apharmaceutically acceptable salt thereof; and, when it is desired toproduce a compound of formula IV where Z is the residue of an estergroup,

(2) esterifying the free carboxyl group at the 3-position of compoundXXII by methods known per se to produce a compound of the formula##STR129## wherein Z is the residue of an ester group and R and R' areas defined above, or a pharmaceutically acceptable salt thereof and, ifdesired, performing one or more of the additional steps of

(d) selectively removing by methods known per se the protecting group R'to produce the desired 4-carboxylic acid compound of formula IV whereinZ is the residue of an ester group;

(e) converting the 4-carboxylic acid compound of step (d) to aphysiologically hydrolyzed ester thereof by methods known per se; or

(f) converting by methods known per se the 4-carboxylic acid compound ofstep (d) or a physiologically hydrolyzed ester thereof to apharmaceutically acceptable salt thereof.

Acyl group R in starting material IIIb may be any organic acyl radicalbut is preferably selected from those acyl groups described above aspreferred in connection with the compounds of general formula III.Carboxyl-protecting group R' may be any easily cleavable ester groupconveniently used to block a free carboxylic acid functional group. Theterm "easily cleavable" has the same meaning as described previously,i.e. removable by methods which do not result in any appreciabledestruction of the remaining portion of the molecule. Preferredprotecting groups are benzhydryl, benzyl, p-nitrobenzyl, trichloroethyl,silyl including especially trimethylsilyl, phenacyl, p-methoxybenzyl,acetonyl, (lower)alkyl including particularly methyl, ethyl and t-butyl,triphenylmethyl, methoxymethyl, acetoxymethyl, phthalidyl, indanyl andpivaloyloxymethyl.

The carboxylation reaction is carries out by reacting thecarboxyl-protected compound IIIb with gaseous carbon dioxide in thepresence of a base. Suitable bases for this step include sodium hydride,n-butyl lithium, t-butyl lithium, lithium dicyclohexylamine, lithium2,2,5,6-tetramethylpiperidine and lithium diisopropylamine. Thepreferred bases are n-butyl lithium and t-butyl lithium. The ester IIIBis dissolved in a dry inert organic solvent, e.g. tetrahydrofuran, anddry gaseous CO₂ introduced into the mixture of base and IIIb solution.The reaction mixture is cooled before introduction of the CO₂ to atemperature in the range of 0° to -80° C. and then allowed to warm toroom temperature with continued addition of carbon dioxide. Uponacidification, e.g. with a mineral acid such as aqueous HCl, compoundXXII may be recovered.

Upon cleavage of carboxyl-protecting group R' to form a free 4-carboxylgroup, e.g. by hydrolysis, chemical reduction or catalytichydrogenation, the 3-carboxymethyl acids prepared by the carboxylationstep or physiologically hydrolyzed esters or pharmaceutically acceptablesalts of said acids or esters may be used as active antibacterialagents.

Compound XXII may also be esterified by known methods to form thedesired carboxyl-protected compound of formula IVa where Z is theresidue of an ester group, most preferably C₁ -C₆ alkyl, e.g. methyl,ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl,n-pentyl, n-heptyl, etc. The ester group Z should be one which isresistant to conditions required for removal of blocking group R', e.g.resistant to hydrogenolysis. The most preferred ester, the methyl ester,may be prepared for example by reaction of compound XXII withdiazomethane. The preferred lower alkyl esters may also be formed by useof the mixed anhydride reaction or by the use of dimethylformamideacetals as described in Helv. Chim. Acta., 48, 1746 (1965). Uponselective cleavage of blocking group R' by known methods, e.g. acidlabile blocking groups such as t-butyl, p-methoxybenzyl ortetrahydropyranyl may be selectively removed by mild acid treatment andgroups such as benzyl or p-nitrobenzyl may be selectively removed byhydrogenolysis, the intermediates of formula IVa where R' is notphysiologically hydrolyzed may be converted to 4-carboxylic acid activeantibacterial agents. The free acid products may optionally be convertedaccording to known methods to physiologically hydrolyzed esters orpharmaceutically acceptable salts thereof.

The compounds and pharmaceutically acceptable salts of formuls XXIIwherein R' is a physiologically hydrolyzed ester group or the de-blockedcompounds of formula XXII or physiologically hydrolyzed esters orpharmaceutically acceptable salts of said acids or esters are asmentioned above active antibacterial agents. These compounds and saltshave approximately the same degree of activity as the acids,physiologically hydrolyzed esters and pharmaceutically acceptable saltsof formula IV, but are more difficult to isolate from the reactionmixture because of the presence of the free carboxyl group on the3-position. For this reason, the preferred compounds and salts offormula IV are those having the esterified carboxymethylene3-substituent, most preferably those having a C₁ -C₆ alkyl radical forthe Z substituent.

The pharmaceutically active compounds of the present invention arepotent antibacterial agents useful in the treatment of infectiousdiseases in poultry and animals, including man, caused by manyGram-positive and Gram-negative bacteria. The active compounds are alsoof value as nutritional supplements in animal feeds and as agents forthe treatment of mastitis in cattle.

The novel medicaments provided by the present invention may beformulated as pharmaceutical compositions comprising, in addition to theactive ingredient, a pharmaceutically acceptable carrier or diluent. Thecompounds may be administered both orally and parenterally. Thepharmaceutical preparations may be in solid form such as capsules,tablets or dragees, or in liquid form such as solutions, suspensions oremulsions. In the treatment of bacterial infections in man, the activecompounds of this invention may be administered parenterally or orallyin an amount of from about 5 to 200 mg/Kg./day and preferably about 5 to20 mg./Kg./day in divided dosage, e.g. three or four times a day. Theyare administered in dosage units containing, for example, 125, 250 or500 mg. of active ingredient with suitable physiologically acceptablecarriers or excipients.

Illustrative examples of the preparation of compounds of the presentinvention follow. These examples ae given in illustration of, but not inlimitation of, the present invention. All temperatures are in degreesCentigrade. AR indicates Analytical Reagent grade. Other abbreviationsinclude TEA for triethylamine, THF for tetrahydrofuran, mm formillimole, TLC for thin layer chromatography and EEDQ is the amide bondforming reagent having the structure ##STR130##

The 7-acylamido compounds prepared in the examples which follow all havethe hydrogen atoms at carbons 6 and 7 cis with respect to each otherand, unless indicated, the products are racemic mixtures in the sensethat they are composed of equal parts of the two isomers having thefollowing structures:

DESCRIPTION OF THE PREFERRED EMBODIMENTS EXAMPLE 1 ##STR132##

Ethyl α-oximinoacetoacetate 1 (80 g.) [Prepared according to the methodof H. Adkins and J. Reeve, JACS, 60, 1328 (1939)] was dissolved in amixture of ethanol (EtOH; 200 ml. USP) and ethanolic HCl (70 ml. of 9.28N HCl-EtOH; 1.25 equiv.). [Amounts of HCl greater and smaller than 1.25equivalents were found to give lower yields of 2.] 10% Palladium oncarbon (8 g.) was added carefully and the mixture was hydrogenated in aParr hydrogenation apparatus starting at 60 psig. After absorption ofthe theoretical amount of hydrogen (1-2 hr.) the catalyst was filteredoff and washed with EtOH. The EtOH was removed in vacuo at 40°-50°leaving a thick red-brown oil. The oil was diluted with 8 vols. ofacetone (AR) with vigorous stirring. Yellow crystals of the aminehydrochloride 2 separated out on cooling, 49 g. (55%), m.p. 122°-123°(cor.) [lit m.p. = 114°-116° uncor.; (W. G. Laver et al., J. Chem. Soc.,(1959), 1474.] [Yields ranged from 45 - 70% according to the scale ofthe reaction.] This material was used without further purification.##STR133##

To a mixture of ethylene glycol (1.75 kg., 28.2 mole) andp-toluenesulfonic acid monohydrate (210 g.; 1.95 mole) which had beenwarmed to 90°, amine hydrochloride 2 (460 g.; 2.54 mole) was added withvigorous mechanical stirring. The mixture was stirred for 40 min. at90°*. The mixture was then poured into a mixture of water (2 l), conc.NH₄ OH (650 ml.), and ice (1 l), and extracted four times with 500 ml.CH₂ Cl₂. The combined CH₂ Cl₂ extracts were washed with brine, dried(Na₂ SO₄), and evaporated to give 491 g. of a dark red oil. The oil wasdiluted to 1.8 1 with Et₂ O (USP), cooled in an ice bath, and EtOH sat'dwith HCl gas was added until the pH reached 2-3. The resulting solid wascollected by filtration and washed with Et₂ O to yield 398 g. 3hydrochloride as a light yellowish solid (70% m.p. 153°-6° (cor.). Ananalytical sample of 3 hydrochloride was recrystallized from2-propanol-Et₂ O, white crystals, m.p. 158°-160° (cor.).

Anal. Calc'd for C₈ H₁₅ NO₄.HCl: C, 42.58; H, 7.15; N, 6.21. Found:42.40; H, 7.24; N, 6.37.

The free base of 3 is conveniently prepared from its hydrochloride bybasification with conc. NH₄ OH and extraction with CH₂ Cl₂. ##STR134##

A mixture of ketal amine 3 (123 g.; 0.65 mole), cinnamaldehyde (85.9 g.;0.65.; 0.65 mole), and CH₂ Cl₂ (3.6 l; AR dried over 4A molecular sieve)was refluxed through a Soxhlet apparatus containing 200 g. of 4Amolecular sieve for 2 hr. The CH₂ Cl₂ was then removed on a rotaryevaporator at 40° with the final traces of CH₂ Cl₂ being removed bypumping down to 0.5 mm. at 25° for 20 minutes. NMR indicated completeformation of imine 4 which was used directly in the next step.

Crude imine 4 (ca. 0.65 mole) was dissolved in CH₂ Cl₂ (3 1; AR driedover 4A molecular sieve), TEA (99.65 ml. 1.1 equiv.) was added, and thesolution was placed under N₂ and cooled to 3°-4° with an ice bath. Asolution of azidoacetyl chloride (85.19 g, 1.1 equiv.) in CH₂ Cl₂ (500ml.) was added with stirring over 4 hr. The reaction mixture was stirredovernight at 25° and then refluxed for 1 hr. The reaction mixture wascooled, washed with 10% HCl, then with brine, and dried over Na₂ SO₄ togive 267 g. of a dark oil which NMR indicated contained 96 weight % of5. (245 g.; 98%).

Anal. sample was recrystallized from methanol, white solid, m.p. 81.5 -82.5° (cor.)

Anal. Calc'd for C₁₉ H₂₂ N₄ O₅ : C, 59.06; H, 5.94: N, 14.50. Found C,59.08; H, 5.73; N, 14.58. ##STR135##

Ethyl ester 5 (64.31 g; 0.168 moles) was dissolved in 700 ml. THF in a 21.3 necked RB flask equipped with magnetic stirring, a thermometer, anadditional funnel and a reflux condenser. There was added 670 ml. of0.25 N NaOH solution (0.168 moles) at such a rate as to hold thetemperature around 25° (took 1 hr.). Stirred at 25° until TLC showedthat 5 had completely reacted (0.75 - 1.25 hr.). The reaction mixturewas carefully acidified to pH 3 with conc. HCl, sat'd with salt, andextracted with CH₂ Cl₂ (3 times). The CH₂ Cl₂ extracts were washed withbrine, dried (Na₂ SO₄) and evaporated in vacuo. The residue wasdissolved in Et₂ O and extracted with 10% NaHCO₃ solution until theextracts were colorless. The combined basic extracts were washed twicewith Et₂ O, then carefully acidified to pH 3 with conc. HCl, sat'd withsalt and extracted with CH₂ Cl₂. The CH₂ Cl₂ extract was washed withbrine, dried (Na₂ SO₄) and evaporated to dryness in vacuo to give 51.84g. (86%) of 6 as a brown solid. Recrystallization from benzene providedan anal. sample as white needles, m.p. 131°-1.5° dec. (cor.).

Anal. Calc'd for C₁₇ H₁₈ N₄ O₅ : C, 56.98; H, 5.06; N, 15.64. Found: C,57.06; H, 5.13; N, 15.78. ##STR136##

Carboxylic acid 6 (128.01 g; 0.358 mole) was dissolved in CH₂ Cl₂ (1 l)and TEA (39.8 g; 0.394 mole) was added [The carboxylic acid 6 referredto was the crystalline solid, m.p. 131-131.5°, which is one of theisomers.]. The mixture was cooled in an icesalt bath to 3° and then amixture of benzyl chloroformate (67.5 g. of 88.6 weight %; 0.394 mole)and CH₂ Cl₂ (200 ml.) was slowly added keeping the reaction temperatureat 3°. After the addition was complete, the reaction mixture was stirredat ambient temperature for 30 min. and then refluxed gently untilevolution of CO₂ ceased (ca. 30 min.). Worked up by washing with 10%HCl, 10% NaHCO₃, brine, dried (Na₂ SO₄) and evaporated to dryness invacuo to yield 161.10 g. (100%) of crude 7. Crystallized frombenzene-petroleum ether (30 - 60) to give 143.60 g. (99%) of 7 as alight beige solid, m.p. 65.5° - 66.5° (cor.).

Anal. calc'd for C₂₄ H₂₄ N₄ O₅ : C, 64.27; H, 5.39; N, 12.49. Found C,64.13; H, 5.36; N, 12.48. ##STR137##

Styryl β-lactam 7 (36.36 g; 81.24 mm) [The styryl β-lactam 7 used inthis experiment was principally one of the possible isomers.] wasdissolved in CH₂ Cl₂ (300 ml.), cooled to -50° to -60° in a dryice-acetone bath, and ozonized until a faint blue color appeared. Thesolution was then flushed with 0₂ until the blue color faded. (CH₃)₂ S(31.87 ml; 5 equiv.) was added to the 31═° solution, which was thenallowed to slowly reach 25° as the cooling bath gradually melted. Keptat 25° under N₂ overnight then washed twice with 1% NaHCO₃, twice withbrine, dried (Na₂ SO₄) and evaporated to dryness. This provided 32.92 g.of 8 an oil which crystallized on standing. This material was slurriedwith ether and filtered to provide 18.84 g. (69%) offwhite solid 8, m.p.97°-100° (cor.). The analytical sample was recrystallized from ether,white crystals, m.p. 101°-102° (cor.).

Anal. Calc'd for C₁₇ H₁₈ N₄ O₆ : C, 54.54; H, 4.84; N, 14.96. Found: C,54.75; H, 4.87; N, 14.89. ##STR138##

Aldehyde 8 (43.15 g; 0.115 mole) was dissolved in THF (400 ml; AR) andthe resultant solution cooled to -5° to -10° (ice-MeOH). NaBH₄ (2.192 g;0.576 mole) was added, with stirring, in three portions. After 25 min.,TLC indicated complete reaction of aldehyde 8. The reaction mixture wascarefully acidified to pH 3 with 10% HCl, diluted with 100 ml. of brine,and extracted with ether (4 ×'s 200 ml.). The combined ether extractswere washed with brine (2 ×'s 150 ml.), dried (Na₂ SO₄), and evaporatedto dryness in vacuo. This provided 48.7 g. of 9 as a red oil (containingsome THF) whose IR and NMR confirmed the structure 9; [ Epimerizationoccurs at the position α to the ester group during this reduction.].This material was used as such in the next reaction.

Crude alcohol 9 (42.37 g; 0.113 mole) and TEA (17.07 ml.; 0.124 mole)were dissolved in CH₂ Cl₂ (400 ml. AR over 4A molecular sieve), placedunder N₂ atmosphere and cooled to 0° to -5° (ice-MeOH). Methanesulfonylchloride (14.15 g, 0.124 mole) dissolved in CH₂ Cl₂ (100 ml, AR) wasadded over 15 minutes. After 1 hr. at ambient temp. the reaction was ca.80% complete (TLC). After 3 hours, the reaction mixture was washed withwater and then with brine, dried over Na₂ SO₄ and evaporated to provide56.1 g. of brown oil. This oil was dissolved in the minimum amount ofbenzene and chromatographed over 700 g. of activated alumina usingEtOAc-Et₂ O (1:3) as eluting solvent. 30 g. of pure crystalline mesylate10 was obtained from the first liter of eluent. 4.2 g. of slightlyimpure mesylate was obtained as a forerun. Total yield = 66% from 8.Anal. sample was crystallized from benzene-ether, white crystals, m.p.97°-99° (cor.).

Anal. Calc'd for C₁₈ H₂₂ N₄ O₈ S: C, 47.61; H, 4.88; N, 12.34. Found: C,47.56; H, 4.93; N, 12.43. ##STR139##

Ketal 10 (3.19 g; 6.43 mm) was placed in a 100 ml. three-necked roundbottom flask equipped with magnetic stirring. 95% TFA (30 ml.) [95% TFAwas prepared by adding 5 ml. of water to a 100 ml. graduated cylinderand diluting to 100 ml. with glacial TFA.] was added and the solutionwas stirred at ambient temp. (ca. 25°) for 2 hr. At this point NMRexamination of an aliquot from the reaction mixture showed completereaction. The mixture was diluted with 10 vols. of brine and extracted 3times with 100 ml. (each) of CH₂ Cl₂. The combined extracts were dried(Na₂ SO₄) and evaporated to dryness in vacuo leaving 3.17 g. of 11 as aheavy brown oil. NMR confirmed the presence of enol 11.

This material was used as such in the next reaction.

Crude enol 11 (described above) (12.02 g; ca. 29.44 mm) was dissolved inCH₂ Cl₂ (100 ml.) and TEA (4.1 ml; 29.44 mm) added. The mixture wasrefluxed under a CaCl₂ drying tube for 2 hr, then washed with 10% HCl,brine and dried (NaSO₄). Evaporation left 8.56 g. of 12 as a light brownoil. This material was taken in CH₂ Cl₂ and filtered through ca. 100 g.of silicic acid powder. Evaporation of the filtrate and Et₂ O washingsgave 6.58 g. of 12 (80.5% from 11) as a light beige solid.

Analytical sample was recrystallized once from ether to give whitecrystals, m.p. 87°-88° (cor.).

Anal. Calc'd for C₁₅ H₁₄ N₄ O₄ : C, 57.32; H, 4.49; N, 17.83. Found: C,57.31; H, 4.58; N, 17.67. ##STR140##

Azido 0-2-isocephem 12 (201 mg; 0.64 mm) was dissolved in absolute EtOH(35 ml.), 87% PtO₂ (100 mg.) was added and the mixture was hydrogenatedat atmospheric pressure. The theoretical amount of hydrogen (for PtO₂reduction) was absorbed in 7 min. and the reaction was stopped to avoidhydrogenolysis of the benzyl ester. The catalyst was filtered off andwashed with 2 vol. of EtOH. Evaporation of the EtOH provided 0.19 g. of13 as a yellowish oil. TLC showed no 12. This oil was used immediatelyin the next step.

Crude amine 13 (0.19 g; 0.64 mm) was dissolved in CH₂ Cl₂ (20 ml.) andphenoxyacetic acid (PAA) (97.4 mg; 0.64 mm) and EEDQ (158 mg; 0.64 mm)were added. The reaction mixture was allowed to stir at 25° for 1 hr.and then it was washed 2 times with 1% NaHCO₃, 2 times with 10% HCl,once with 3 vols. of brine, dried (Na₂ SO₄) and evaporated to dryness invacuo. This provided 0.18 g. of 14 as a yellowish gum which wastriturated with dry Et₂ O. The Et₂ O triturant was cooled at 0°overnight. The resultant crystals were collected by filtration andwashed once with petroleum ether (30° - 60°) to give white crystals of14, m.p. 133°-135° dec. (cor.).

Anal Calc'd for C₂₃ H₂₂ N₂ O₆ : C, 65.39; H, 5.25; N, 6.63. Found: C,65.22; H, 5.31; N, 6.86. ##STR141##

Benzyl ester 14 (100 mg; 0.237 mm) was dissolved in a mixture ofabsolute EtOH (10 ml.) and THF (7 ml.). 10% Pd-C (100 mg.) was carefullyadded and the mixture was hydrogenated at atmospheric pressure. Hydrogenuptake was complete after ca. 7 min. The catalyst was filtered off andwashed once with EtOH. The EtOH was removed in vacuo leaving 90 mg. ofpartly crystalline residue. The residue was crystallized fromacetone-ether to provide off-white crystals of 15, m.p. 171°-172° dec.(cor.).

Anal. Calc'd for C₁₆ H₁₆ N₂ O₆ : C, 57.83; H, 4.85; N, 8.43. Found: C,57.67; H, 4.97; N, 8.34.

A sample of compound 15 prepared above which can be named7β-phenoxyacetamido-3-methyl-O-2-isocephem-4-carboxylic acid (calledBC-L8) after solution in water and dilution with Nutrient Broth wasfound to exhibit the following Minimum Inhibitory Concentrations(M.I.C.) in mcg./ml. versus the indicated miroorganisms as determined byovernight incubation at 37° C. by Tube dilution. One old, orallyabsorbed cephalosporin (cephalexin) was included.

                  Table 1                                                         ______________________________________                                        M.I.C. in mcg./ml.                                                                                          Cepha-                                          Organism            BC-L8     lexin                                           ______________________________________                                        D. pneumoniae A9585     .5        .6                                          +5% serum*                                                                    Str. Pyogenes A9604     .5        .6                                          +5% serum*                                                                    S. aureus Smith.sup.+                                                                       A9537     .5        1.3                                         S. aureus Smith.sup.+                                                                       A9537     2         2.5                                         +50% serum                                                                    S. aureus BX1633-2                                                                          A9606     2         1                                           at 10.sup.-3 dil'n                                                            S. aureus BX1633-2                                                                          A9606     32        2                                           at 10.sup.-2 dil'n                                                            Sal. enteritidis.sup.+                                                                      A9531     8         2                                           E. coli Juhl.sup.+                                                                          A15119    63        4                                           E. coli.sup.+ A9675     250       16                                          K. pneumoniae.sup.+                                                                         A9977     32        4                                           K. pneumoniae.sup.+                                                                         A15130    500       16                                          Pr. mirabilis.sup.+                                                                         A9900     16        4                                           Pr. morganii.sup.+                                                                          A15153    500       >125                                        Ps. aeruginosa.sup.+                                                                        A9843A    500       >125                                        Ser. marcescens.sup.+                                                                       A20019    500       >125                                        Ent. cloacae  A9656     500       >125                                        Ent. cloacae  A9657     63        2                                           Ent. cloacae  A9659     500       125                                         S. aureus meth.-                                                                            A15097    32        16                                          resist; at 10.sup. -3                                                         dil'n                                                                         ______________________________________                                         *50% Nutrient Broth - 45% Antibiotic Assay Broth                              .sup.+ at 10.sup.-4 dilution.                                            

EXAMPLE 2 ##STR142## Benzyl Oximino-Acetoacetate

The procedure was essentially the same as that described for thecorresponding ethyl ester by H. Adkins and J. Reeve, JACS 60, 1328(1938).

In a three necked one liter flask, fitted with a thermometer, a droppingfunnel and a magnetic stirrer were placed 173 g. (0.9 mole) of benzylacetoacetate [The benzyl acetoacetate was prepared as described by Bakeret al., J. Org. Chem. 17, 91 (1952)] and 130 ml. of glacial acetic acid.The contents were cooled in an ice-salt bath and a solution of 69 g. (1mole) of sodium nitrite in 130 ml. of water was added over a period ofhalf an hour; the temperature was kept at 0° to 10° C. After thereaction mixture was stirred for one hour at room temperature, 400 ml.of water was added and the stirring was continued for an additional twohours. The reaction mixture was extracted three times with 200 ml.portions of diethyl ether. The diethyl ether extracts were combined,washed once with water, three times with saturated sodium bicarbonatesolution and once with brine. After drying over anhydrous sodiumsulfate, the diethyl ether solution was evaporated leaving 16 as a clearoil which solidified upon trituration with petroleum ether (30°- 60°) togive 186.5 g. (93.2%) of white solid. Its NMR spectrum was consistentwith the assigned structure. Generally the product was used as such insubsequent reaction but it can be recrystallized from toluene, m.p. 81°-82° C. ##STR143##

Benzyl Oximino-Acetoacetate Ethylene Ketal

In a two liter flask fitted with a Dean Stark water separator and acondenser were placed 186.5 g. (0.85 mole) of benzyloximino-acetoacetate (17), 62 g. (1 mole) of ethylene glycol, 800 ml. ofbenzene (reagent grade) and 2 g. of toluenesulfonic acid. The reactionmixture was boiled at reflux until 15 ml. of water was removed (3hours). The benzene solution was washed once with saturated sodiumbicarbonate solution and once with brine. After drying over anhydroussodium sulfate, the benzene solution was evaporated, leaving 212 g.(94%) of 17 as a light yellow oil. Its NMR spectrum was consistent withthe assigned structure. Generally the compound was used as used insubsequent reaction but one of the isomers can be crystallized in 35%yield from toluene-petroleum ether, m.p. 52° C. ##STR144##

Benzyl Amino-Acetoacetate Ethylene Ketal (18)

[The procedure was essentially the same as that described for thereduction of unsaturated hydroxyimino ethyl esters by D. J. Drinkwaterand P. W. G. Smith, J. Chem. Soc. (C), 1305 (1971).] [Aluminum amalgamwas prepared essentially the same as that described in "Vogel" exceptthe following modifications:

A. 5% NaOH was used.

B. The second washing with ethanol was omitted.

C. Dry diethyl ether was used for washing and most of the water must bedrained.]

[Vogel "Practical Organic Chemistry" 3rd Edition. Longemans Green & Co.,London (1957) p. 198.]

Aluminum amalgam (from 27 g. of aluminum foil) freshly prepared in athree-necked one liter flask was covered with 500 ml. of diethyl ether.The flask was fitted with a mechanical stirrer, a condenser, and adropping funnel. 132.5 g. (0.5 mole) of benzyl oximino-acetoacetateethylene ketal (17) in 300 ml. of wet diethyl ether was added dropwiseat such a rate as to maintain boiling at reflux. After stirring for fourhours, the reaction mixture was filtered through a Buchner funnel. Thefiltrate was evaporated leaving 110 g. of 18 as a yellowish oil. The oilwas picked up in 800 ml. of dry diethyl ether and dry hydrogen chloridewas passed in to give 108 g. of white hydrochloride salt of 18 which wascollected, m.p. 157°-158° C.

To obtain the free base 18, the hydrochloride salt was suspended in 500ml. of diethyl ether, concentrated ammonium hydroxide was added withshaking until most of the solid went into solution, then washed twicewith brine. After drying over anhydrous magnesium or sodium sulfate, thesolvent was evaporated leaving 90 g. of 18 as a colorless oil (71%). ItsNMR spectrum was consistent with the assigned structure. ##STR145##

Schiff Base Formation

In a one liter flask fitted with a Dean Stark water separator and acondenser were placed 70.3 g. (0.28 mole) benzyl aminoacetoacetateethylene ketal (18), 37 g. (0.28 mole) cinnamaldehyde and 750 ml. ofmethylene chloride. The mixture was boiled at reflux for half an hourand then 400 ml. of methylene chloride was distilled and removed throughthe Dean Stark water separator. The concentrated solution was firstdried over anhydrous sodium sulfate and then evaporated completely on anevaporator to drive the reaction to completion. The residual, oily 19was checked by NMR to ensure complete Schiff base formation beforecontinuing on the next step.

β-Lactam Formation

The freshly prepared Schiff base (19) was diluted with 600 ml. ofmethylene chloride and cooled to 0°0 C. (ice-salt bath). [All themethylene chloride used in the cyclo addition reaction was reagent gradewhich was first dried over molecular sieve (Type 4A) and then overanhydrous calcium chloride.] 31.1 g. (0.308 mole) of triethylamine wasadded and followed by a solution of 36.2 g. (0.308 mole) of azido-acetylchloride in 362 ml. of methylene chloride adding dropwise at 0° C. overa period of one hour. The reaction mixture was stirred for an additionalhour at room temperature and then evaporated on a flash evaporator atreduced pressure while being heated on a 35° C. water bath; thisoperation is necessary to ensure complete β-lactam formation. Theresidue was diluted with 500 ml. of diethyl ether and filtered. Thefiltrate was washed twice with brine and dried over anhydrous sodiumsulfate. Evaporation of this solution yielded 117.5 g. (94%) of product20. Its NMR and IR spectra were consistent with the assigned structure.Generally, the product was used as such in subseqent reaction but one ofthe isomers can be crystallized from diethyl ether.

Compound 20 is identical with compound 7 prepared in Example 1 and isreacted according to the procedures of Example 1 to produce benzyl7β-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylate having formula##STR146##

A mixture of amine 13 (430 mg.; 1.49 mmole),N-carbobenzoxy-D(-)-phenylglycine (422 mg.; 1.49 mmole), EEDQ (368 mg.;1.49 mmole) and 30 ml. CH₂ Cl₂ was stirred at 25° for 1 hour. It wasthen washed with 10% HCl, 1% NaHCO₃ and brine and dried over Na₂ SO₄.Evaporation in vacuo gave 0.85 g. of a white foam which was crystallizedfrom ether-pentane and then from MEOH.

Anal. Calc'd for C₃₁ H₂₉ N₃ O₇.0.5 H₂ O: C, 65.95; H, 5.36; N, 7.44.Found: C, 66.12; H, 5.30; N, 7.57.

The white foam was characterized by IR and NMR to be the N-protectedcompound of formula ##STR147##

A mixture of the above N-protected intermediate (283 mg.), 600 mg. of30% Pd-diatomaceous earth and 10 ml. of ethanol was hydrogenated at 25°at 50 psig in a Parr hydrogenator. After 0.5 hour, the catalyst wasfiltered off and the filtrate was evaporated to dryness in vacuo. Theresultant residue was covered with CHCl₃ and then HCl gas wasintroduced. Addition of ether produced a precipitate which was filteredoff and dried in vacuo. The product, a yellowish solid, decomposed at177°-182° (corrected) and was characterized by IR and NMR as7β-(D-α-aminophenylacetamido)-3-methyl-Δ³ -O-2-isocephem-4-carboxylicacid hydrochloride (called BC-L9) of the formula ##STR148##

Anal Calc'd for C₁₆ H₁₇ N₃ O₅. HCl.H₂ 0: C, 49.81; H, 5.22; N, 10.89.Found: C, 49.76; H, 5.21; N, 9.11.

A sample of BC-L9 after solution in water and dilution with NutrientBroth was found to exhibit the following Minimum InhibitoryConcentrations (M.I.C.) in mcg./ml. versus the indicated microorganismsas determined by overnight incubation at 37° C. by tube dilution. Oneold, orally absorbed cephalosporin (cephalexin) was included.

                  Table 2                                                         ______________________________________                                        M.I.C. in mcg./ml.                                                                                          Cepha-                                          Organism            BC-L9     lexin                                           ______________________________________                                        D. pneumoniae A9585     .5        .3                                          +5% serum*                                                                    Str. pyogenes A9604     .5        .3                                          +5% serum*                                                                    S. aureus Smith.sup.+                                                                       A9537     2         1.3                                         S. aureus Smith.sup.+                                                                       A9537     16        2.5                                         +50% serum                                                                    S. aureus BX1633-2                                                                          A9606     8         1                                           at 10.sup.-3 dil'n                                                            S. aureus BX1633-2                                                                          A9606     32        2                                           at 10.sup.-2 dil'n                                                            S. aureus meth.-                                                                            A15097    32        16                                          resist; at 10.sup.-3                                                          dil'n                                                                         Sal. enteritidis.sup.+                                                                      A9531     16        4                                           E. coli Juhl.sup.+                                                                          A15119    32        4                                           E. coli.sup.+ A9675     63        8                                           K. pneumoniae.sup.+                                                                         A9977     16        4                                           K. pneumoniae.sup.+                                                                         A15130    63        16                                          Pr. mirabilis.sup.+                                                                         A9900     63        8                                           Pr. morganii.sup.+                                                                          A15153    250       >125                                        Ps. aeruginosa.sup.+                                                                        A9843A    >500      >125                                        Ser. marcescens.sup.+                                                                       A20019    500       >125                                        Ent. cloacae  A9656     500       >125                                        Ent. cloacae  A9657     32        4                                            Ent. cloacae A9659     125       >125                                        ______________________________________                                         *50% Nutrient Broth - 45% Antibiotic Assay Broth                              .sup.+ at 10.sup.-4 dilution.                                            

EXAMPLE 3 7β-Amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylic acid##STR149##

A mixture of benzyl 7β-azido-3-methyl-Δ³ -O-2-isocephem-4-carboxylate(314 mg.; 1 mmole), 30% Pd-diatomaceous earth (274 mg.) and 25 ml. ofabsolute EtOH was hydrogenated at 25° and atmospheric pressure. Hydrogenuptake was complete after 20 minutes. The reaction mixture was thenwarmed to 40°, the catalyst was filtered off and washed with 1 volume ofEtOH. The combined filtrate and washings were evaporated to dryness invacuo to provide 140 mg. (71%) of white powdery residue which wasidentified by IR and NMR as the title product. Decomp. >ca. 209° (cor).

Anal. Calc'd for C₈ H₁₀ N₂ O₄.0.5H₂ O: C, 46.38; H, 5.35; N, 13.52.Found: C, 46.86; H, 5.35; N, 13.58.

A sample of the above compound (called BC-L61) was found to inhibit S.aureus A9537 at a concentration of >125 mcg./ml., E. coli A15119 at aconcentration of >125 mcg./ml., D. pneumoniae A9585 at a concentrationof >8 mcg./ml. and St. pyrogenes at a concentration of >8 mcg./ml.

EXAMPLE 4 Benzyl 7β-Amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylate##STR150##

H₂ S gas was bubbled into a solution of 2.0 g. (6.35 mmoles) ofbenzyl-7-β-azido-3-methyl-Δ³ -O-2-isocephem-4-carboxylate (12), 4.2 ml.(30 mmole) triethylamine, and 50 ml. of AR methylene chloride containedin a 100 ml. 3 necked flask equipped with a condenser, a gas inlet tubeand magnetic stirring. The solution color changed from colorless toorange and a gas (N₂) is evolved. TLC after 20 minutes showed that thereaction was complete. The reaction mixture was evaporated to drynessand the resulting yellow residue was shaken three times with a 1:1mixture of 10% aqueous HCl and ether. Almost all of the residue goesinto solution. The aqueous layer was separated and the ether layer(yellow colored) was washed with 10% aqueous HCl. The combined HClphases were washed once with ether and then carefully alkalized withsolid NaHCO₃. The alkalized aqueous phase was extracted twice with CH₂Cl₂. The CH₂ Cl₂ extracts were combined, washed twice with saturatedbrine, dried (Na₂ So₄) and evaporated to dryness in vacuo. This provided1.25 g (68%) of a gummy residue (some black impurities) whichcrystallized upon standing overnite. NMR on this material indicates apurity of at least 95%. The solid could be recrystallized from ether togive a white solid, mp. 91°-92° (cor).

Anal. Calc'd for C₁₅ H₁₆ N₂ O₄ : C, 62.49; H, 5.59; N, 9.72. Found: C,62.54; H, 5.51; N, 9.65.

EXAMPLE 5 7β-Phenoxyacetamido-3-benzyl-Δ³ -O-2-isocephem-4-carboxylicacid ##STR151##

The ethyl γ-phenylacetoacetate was prepared after the proceduredescribed¹ in 40% yield. B.P. 0.005 mm 103°-105°.

6 1. G. R. Ames and W. Davey. J. Chem. Soc. 1957, 3480-87.

    φCH.sub.2 COCH.sub.2 COOEt+ φCH.sub.2 OH → φCH.sub.2 COCH.sub.2 COOBz

A mixture of ethyl γ-phenylacetoacetate (166 g.; 0.76 mole) and benzylalcohol (100 g.; 0.92 mole) was immersed into an oil bath, preheated to170°, and with stirring the ethyl alcohol produced was distilled off.With an aspirator, a forerun fraction (B.P. 100 mm 65°-80°) was removedand finally the residue was distilled at low pressure. B.P. 0.002 mm155°-157°. By recycling the forerun, a further quantity of pure compoundwas obtained. Overall yield was 171 g. (84%). ##STR152##

A solution of benzyl γ-phenylacetoacetate 54 (85.5 g.; 0.32 mole) inglacial acetic acid (400 ml.) and water (150 ml.) was cooled to 5° in anice bath and while stirring vigorously, to it was added dropwise in 4minutes a solution of sodium nitrate (25.5 g.; 0.37 mole) in water (100ml.). The reaction temperature rose to 16° C. and was stirred withcooling for 30 minutes more. The cooling bath was removed and stirringcontinued for 2 hours. 800 ml. of water was added and the solution wasextracted with 3 × 100 ml. CH₂ Cl₂. The combined extracts were washedwith water and brine, dried over Na₂ SO₄ and evaporated, leaving 92.50g. oil. This was crystallized in 90 ml. CCl₄ to give light yellowcrystals 52.80 g. (56%), m.p. 69°-70° C. ##STR153##

A mixture of oxime 55(5.94 g.; 20 mm), ethylene glycol (1.36 g.; 22 mm)and p-toluenesulfonic acid monohydrate (0.59 g.) in benzene (100 ml.;A.R.) was refluxed over a Dean-Stark water trap for 31/2 hours. It wascooled, and poured into 100 ml. saturated NaHC0₃ and extracted withbenzene. After the organic phase was washed with water, dried over Na₂SO₄ and evaporated, 6.70 g. of an oil was obtained. It was crystallizedin 20 ml., CCl₄, and gave 4.0 g. (57%) light yellow solid, m.p. 90°-92°C. ##STR154##

Aluminium foil (6.9 g.; 255 mm) cut into small strips and looselyfolded, was covered with 5% NaOH and allowed to react for 2 minutes. Itwas decanted and washed successively with water and 95% EtOH, thencovered with 2% mercuric chloride, and allowed to react for 2 minutes.After decanting, it was washed with water and ether, then covered with"wet ether". To this amalgam with stirring was added an ether solutionof the "oximinioester", (29.5 g.; 85 mm) in ether (600 ml.). There was amildly exothermic reaction and after the addition was complete, it wasrefluxed for 2 hours. It was cooled, filtered through celite andextracted with 4 × 100 ml. 10% HCl. White crystals separated from theaqueous phase and were filtered, washed with cold water and dried togive 29.0 g. solid, m.p. 181°-183° with decomposition. Recrystallizedfrom EtOH/ether, m.p. 182°-184° with decomposition. The free base wasobtained by suspending the hydrochloride in water and alkalizing withcold concentrated NH₄ OH. Yield 100%.

Anal. Calc'd. for C₁₉ H₂₂ NO₄.HCl: C, 62.72; H, 6.09; N, 3.85. Found: C,62.83; H, 6.14; N, 3.84. ##STR155##

Ketal-amine 57(56.70 g.; 0.173 mole) was dissolved in dry Ch₂ Cl₂ (600ml.) and to it was added cinnamaldehyde (23.0 g.; 0.173 mole). Thesolution was refluxed for 30 minutes and the solvent was removed on theaspirator. The residue was redissolved in CH₂ Cl₂ (600 ml.), the flaskwas fitted with a Dean-Stark water trap, and the solvent refluxed while300 ml. of CH₂ Cl₂ was collected and continuously removed through thetrap. The residue was dried over Na₂ SO₄ and evaporated to drynessleaving 77.32 g. light yellow oil. This was redissolved in CH₂ Cl₂ (300ml.). Triethylamine (27 ml.; 0.19 mole) was added and while stirring andcooling at 3°-5° C. in an ice-bath there was added dropwise a solutionof azidoacetyl chloride (22.8 g.; 0.19 mole) in CH₂ Cl₂ (300 ml.),addition being done over 2 hours. It was kept at room temperature undernitrogen for 16 hours, and refluxed for 1 hour. The solution was cooled,washed with 10% HCl, then with brine, dried over Na₂ SO₄ and evaporatedto give 90.35 g. It was used as such with no further purification.##STR156##

Styryl β-lactam 59(11.0 g.; 21 mm) was dissolved in CH₂ Cl₂ (150 ml.)and ozonized at -60° until a blue color appeared, then O₂ was used toflush away the excess ozone. To the solution was added 7.7 ml. (105mmole) of dimethyl sulfide and the cooling bath was allowed to warm upto 25° C. spontaneously in 5 hours. The solution was kept at 25° for 16hours then washed with 1% NaHCO₃ and brine, dried over Na₂ SO₄ andevaporated to dryness. It was triturated with petroleum ether (30°-60°)and the residue kept at 0.05 mm/Hg and 45° C for 16 hours. This removedall the benzaldehyde, and left 8.07 g. oil. It was used as such withoutany further purification. ##STR157##

A solution of aldehyde 60(2.1 g.; 5 mm) in THF (50 ml.; A.R.) was cooledto -5° in an ice-salt bath and while stirring, to it was added sodiumborohydride (0.1 g.; 2.9 mm), all at once. It was stirred at 0°-5° for30 minutes, then the cooling bath was removed and stirring continued atroom temperature for 30 minutes. It was carefully acidified with cold10% HCl, saturated with sodium chloride and extracted with ether. Theether phase was washed with water, and brine, dried over Na₂ SO₄ andevaporated to give 1.88 g. oil. It was purified by chromatography onsilica gel III, eluting with ether/petroleum ether 2:1. ##STR158##

A mixture of alcohol 61 (1.10 g.; 2.6 mm) and triethylamine (0.29 g.;2.85 mm) in CH₂ Cl₂ (25 ml.) was cooled to 0° C. and with stirring undernitrogen to it was added dropwise a solution of methanesulfonyl chloride(0.33 g.; 2.85 mm) in CH₂ Cl₂ (10 ml.). It was stirred at 0° for 15minutes, then at room temperature for 1 hour. It was washed with waterthen with brine, dried over Na₂ SO₄ and evaporated to give 1.36 g. oil.It was used as such with no further purification. ##STR159##

The ketal-mesylate 62 (1.36 g.; 2.5 mm) was dissolved in 95%trifluoroacetic acid (15 ml.) and stirred at 50°-55° for 2 hours on anoil bath. It was poured into brine and extracted with CH₂ Cl₂. Afterwashing the organic extracts with water and drying over Na₂ SO₄, thesolvent was removed on the aspirator and left 1.20 g. red oil. Nofurther purification was attempted. ##STR160##

A mixture of crude enol-mesylate 63 (5.4 g.) and triethylamine (2 ml.)in dry CH₂ Cl₂ (100 ml. was refluxed for 5 hours. It was cooled, washedwith 10% HCl and water, dried over Na₂ SO₄ and evaporated on theaspirator to give 4.24 g. oil. This was purified by chromatography on200 g. of silica gel III, eluting with ether/petroleum ether 2:1. Thepure compound 64 crystallized, m.p. 117°-118°. ##STR161##

A mixture of "azido-isocephem" 64 (0.49 g.; 1.25mm) and triethylamine(0.9 ml.; 6.5 mm) in CH₂ Cl₂ (50 ml.) was cooled in an ice bath andwhile being stirred, was saturated with H₂ S. The cooling bath wasremoved and there was gas evolution which subsided in 10 minutes. Atthis point, T.L.C. showed no starting material remained. Attempts toextract the amine from the solution as its hydrochloride failed as it ismore soluble in CH₂ Cl₂ than in water. The CH₂ Cl₂ solution of the freebase was dried over Na₂ SO₄ and evaporated on the aspirator to leave0.40 g. of a semi-solid. It was used as such with no furtherpurification. ##STR162##

A solution of "amine-isocephem" 65 (0.48 g.; 1.25 mm), phenoxyaceticacid (0.19; 1.25 mm) and EEDQ (0.31 g.; 1.25 mm) in CH₂ Cl₂ (100 ml.)was stirred at room temperature for 16 hours. It was washed with 1%NaHCO₃ solution, then with brine, dried over Na₂ SO₄ and evaporated onthe aspirator to leave 0.56 g. of a slightly yellow gum. It was washedwith no further purification. ##STR163##

The "isocephem compound"66 (0.49 g.; 1 mm) was dissolved in ethylacetate (100 ml.) and glacial acetic acid (10 ml.), 20% Pd(OH)₂ oncarbon (0.50 g.) was added and the mixture was agitated on a Paarapparatus at 60 psi of H₂ for 2 hours. The solid was filtered off oncelite and the filtrate evaporated to dryness. The residue was extractedwith saturated NaHCO₃, the aqueous phase was acidified with 10% HCl andextracted with CH₂ Cl₂. This was then washed with water, dried over Na₂SO₄ and evaporated to dryness. The resulting solid was recrystallizedfrom benzene and gave white crystals, m.p. 123°-125° with decomposition.

Anal. Calc'd. for C₂₂ H₂₀ N₂ O₆ : C, 64.70; H, 4.94; N, 6.86. Found: C,64.78; H, 4.87; N, 6.80.

A sample of compound 67 prepared above which can be named7β-phenoxyacetamido-3-benzyl-Δ³ -O-2-isocephem-4-carboxylic acid (alsocalled BC-L17) after solution in water and dilution with Nutrient Brothwas found to exhibit the following Minimum Inhibitory Concentrations(M.I.C.) in mcg./ml. versus the indicated microorganisms as determinedby overnight incubation at 37° C. by tube dilution. Cephalexin wasincluded as a comparison compound.

                  Table 3                                                         ______________________________________                                        M.I.C. in mcg./ml.                                                                                          Cepha-                                          Organism            BC-L17    lexin                                           ______________________________________                                        D. pneumoniae A9585     .25       .16                                         +5% serum*                                                                    Str. pyogenes A9604     .25       .16                                         +5% serum*                                                                    S. aureus Smith.sup.+                                                                       A9537     .25       .6                                          S. aureus Smith.sup.+                                                                       A9537     1         1.3                                         +50% serum                                                                    S. aureus BX1633-2                                                                          A9606     8         1                                           at 10.sup.-3 dil'n                                                            S. aureus BX1633-2                                                                          A9606     >125      2                                           at 10.sup.-2 dil'n                                                            S. aureus meth.-                                                                            A15097    125       16                                          resist; at 10.sup.-3                                                          dil'n                                                                         Sal. enteritidis.sup.+                                                                      A9531     32        2                                           E. coli Juhl.sup.+                                                                          A15119    >125      8                                           E. coli.sup.+ A9675     >125      16                                          K. pneumoniae.sup.+                                                                         A9977     >125      4                                           K. pneumoniae.sup.+                                                                         A15130    >125      16                                          Pr. mirabilis.sup.+                                                                         A9900     >125      4                                           Pr. morganii.sup.+                                                                          A15153    >125      >125                                        Ps. aeruginosa.sup.+                                                                        A9843A    >125      >125                                        Ser. marcescens.sup.+                                                                       A20019    >125      >125                                        Ent. cloacae  A9656     > 125     >125                                        Ent. cloacae  A9657     >125      4                                           Ent. cloacae  A9659     >125      >125                                        ______________________________________                                         *50% Nutrient Broth - 45% Antibiotic Assay Broth                              .sup.+ at 10.sup.-4 dilution.                                            

EXAMPLE 6 7β-Phenoxyacetamido-b 3-phenethyl-Δ³-O-2-isocephem-4-carboxylic acid ##STR164##

Phenylmagnesium bromide was prepared¹ in the usual way from magnesium(24.30 g.; 1 mole), phenethylbromide (204 g.; 1.1 mole) and a trace ofiodine in ether (250 ml.; AR). While maintaining the reactiontemperature at 25°-30° C., to it was added ethylcyanoacetate (45.2 g.;0.25 mole) and the resulting solution was stirred at room temperaturefor 24 hours. It was decomposed with saturated ammonium chloride, 10%HCl and water and the phases separated. The organic extract was treatedwith 10% HCl (250 ml.) and stirred vigorously for 3 hours. The organicphase was washed with brine, dried over Na₂ SO₄ and evaporated on theaspirator to give 83.5 g. oil. Fractional distillation gave 12.67 g.(21%) pure product. B.P. 0.1 millimeter 114°-122° C.

A mixture of ethyl γ-benzylacetoacetate (12.56 g.; 50 mm) and benzylalcohol (8.1 g.; 75 mm) was immersed into an oil bath preheated to 170°C. and stirred. After ethanol had been filtered off, the excess benzylalcohol was removed on the aspirator and finally the residue wasdistilled. B.P. 0.01 millimeter, 160°-163°. 6.15 g. pure product wasobtained. ##STR166##

To a solution of benzyl γ-benzylacetoacetate (37.0 g.; 0.13 mole) inglacial acetic acid (50 ml.) was added dropwise in 1 hour a solution ofsodium nitrite (10 g.; 0.143 mole) in water (50 ml.). The reactiontemperature was kept at 24°-30° C. It was stirred an additional hourafter the addition then diluted with water (100 ml.), and extracted withether. The organic extract was washed with water and with 1% NaHCO₃until the washings were basic, then with brine. It was then dried overNa₂ SO₄ and evaporated on the aspirator to leave 40.1 g. of a lightyellow oil. It was used as such with no further purification. ##STR167##

A mixture of oxime (8.48 g.; 27 mm), ethylene glycol (1.85 g.; 30 mm)and p-toluenesulfonic acid monohydrate (0.85 g.) in benzene (100 ml.;AR) was refluxed over a Dean-Stark water trap for 4 hours. It wascooled, poured onto saturated NaHCO₃ and after shaking well, the phaseswere separated. The organic phase was washed with water and brine, driedover Na₂ SO₄ and evaporated on the aspirator to leave 9.4 g. of an oil.It was used without further purification. ##STR168##

Aluminum foil (27 g.; 1 mole) cut into small strips and loosely foldedwas covered with 5% NaOH and allowed to react for 2-3 minutes. It wasdecanted and washed successively with water and 95% EtOH, then coveredwith 2% mercuric chloride and allowed to react for 2 minutes. Afterdecanting, it was washed with water and ether, and finally covered with"wet water". To this amalgam with stirring was added an ether solutionof the "oximinoester" 70 (43 g.; 0.12 mole) in ether (300 ml.). Therewas an exothermic reaction and after it subsided, the system wasrefluxed for 4 hours. The inorganic material was filtered on celite andthe filtrate shaken well with 10% HCl (100 ml.) White crystals separatedand were collected by filtration, washed with ether and dried in adessicator to give 54.0 g. solid, m.p. 186°-188°. The free base wasobtained by suspending the solid in water, carefully alkalizing withcold concentrated Na₄ OH and extracting with CH₂ Cl₂. After evaporationof the solvent, 27.91 g. of a yellow oil was obtained. ##STR169##

A mixture of "amine-ketal" 71 (27.6 g.; 81 mm) and cinnamaldehyde (10.7g.; 81 mm) in CH₂ Cl₂ (500 ml.) was refluxed over a Dean-Start watertrap while 200 ml. solvent was removed. By replacing the solvent removedwith fresh one, a further 3 × 250 ml. fractions were removed also.Finally, the residue was evaporated to dryness on the aspirator andpumped down at 0.05 millimeter/Hg. and 40° C. for 10 minutes. Theresidue was re-dissolved in CH₂ Cl₂ (250 ml.), triethylamine (11.9 ml.;85 mm) was added and while stirring and cooling in an ice bath, to itwas added dropwise in 2 1/2 hours a solution of azidoacetyl chloride(10.15 g.; 85 mm) in CH₂ Cl₂ (100 ml.). It was kept at room temperatureunder nitrogen for 16 hours and refluxed for 1 hour. The solution wascooled, washed with 10% HCl, then with brine, dried over Na₂ SO₄ andevaporated on the aspirator to give 44.77 g. red oil. It was used assuch with no further purification. ##STR170##

Styryl β-lactam 73(6.7 g.; 12.5 mm) was dissolved in CH₂ Cl₂ (150 ml.),and ozonized at -78° until a blue color appeared, and then nitrogen wasused to flush away the excess ozone. To the solution was added dimethylsulfide (4.5 ml.; 65 mm) and the cooling bath was allowed to warm up to25° spontaneously. The solution was kept at room temperature for 16hours then washed with 1% NaHCO₃ and brine dried over Na₂ SO₄ andevaporated to dryness. It was then pumped down at 0.05 millimeter/Hg.and 50° C. for 20 hours to remove most of the benzaldehyde. The residuewas chromatographed on 250 g. of silica gel III, eluting with; firstether/petroleum ether 2:1 (to remove benzaldehyde) and then with ether.##STR171##

A solution of aldehyde 74(4.1 g.; 8.8 mm) in THF (100 ml.; A.R.) wascooled to -5° and while stirring, to it was added sodium borohydride(0.17 g.; 4.5mm) all at once. It was stirred at -5° C. for 1 1/2 hours,then carefully acidified with 10% HCl, saturated with sodium chlorideand extracted with ether. The ether phase was washed with water, andbrine, dried over Na₂ SO₄ and evaporated to give 3.8 g. oil. It was usedwith no further purification. ##STR172##

A mixture of alcohol 75 (3.8 g.; 8.3 mm) and triethylamine (1.25 ml.; 9mm) in CH₂ Cl₂ (50 ml.) was cooled to 0°, and with stirring undernitrogen, to it was added dropwise, a solution of methanesulfonylchloride (1.0 g.; 9 mm) in CH₂ Cl₂ (25 ml.). It was stirred at roomtemperature for 1 1/2 hours, then washed with 10% HCl water and brine,and dried over Na₂ SO₄. It was then evaporated to give 4.3 g. oil. Thiswas purified by chromatography on 250 g. silica gel III eluting withether/petroleum ether 3:1. 2.05 g. of pure mesylate was obtained.##STR173##

Ketal-mesylate 76(2.05 g.; 3.7 mm) was dissolved in 95% trifluoroaceticacid (200 ml.) and stirred at 50°-55° C. for 2 hours on an oil bath. Itwas then poured into a mixture of crushed ice and brine and extractedwith CH₂ Cl₂. After washing the organic extracts with water, and dryingover Na₂ SO₄, the solvent was removed on the aspirator and left 1.73 g.oil. No further purification was attempted. ##STR174##

A mixture of crude "enol-mesylate" 78 (1.71 g.; 3.4 mm) andtriethylamine (0.48 ml.; 3.4 mm) in CH₂ Cl₂ (50 ml.) was refluxed for 5hours. It was cooled, washed with 10% HCl and water, dried over Na₂ SO₄and evaporated on the aspirator to give 1.35 g. oil. This was purifiedby chromotography on 75 g. silica gel III eluting with ether/petroleumether 2:1. The pure cis-β-lactam was obtained as white crystals, m.p.97°-98° (MeOH).

Anal. Calc'd. for C₂₂ H₂₀ N₄ O₄ : C, 65.34; H, 4.98; N, 13.85. Found: C,65.36; H, 4.96; N, 13.97. ##STR175##

A mixture of "azido isocephem" 80, (0.81 g.; 2 mm) and triethylamine(0.56 ml.; 4 mm) in CH₂ Cl₂ (50 ml.) was cooled in an ice bath and whilebeing stirred, was saturated with H₂ S. The cooling bath was removed andthere was gas evolution. After stirring at room temperature for 1 hour,the solution was evaporated at room temperature and partitioned betweenether and 10% HCl. White crystals separated and were collected byfiltration, washed with ether and dried to give 1.12 g. white solid,m.p. 120°-123° with decomposition. The free base was obtained bysuspending the solid in water, alkalizing with cold concentrated NH₄ OHand extracting with CH₂ Cl₂. This was washed with brine, dried over Na₂SO₄ and evaporated on the aspirator. ##STR176##

A solution of "amino isocephem" 80 (0.49 g.; 1.05 mmole), phenoxyaceticacid (0.16 g.; 1.05 mm) and EEDQ (0.26 g.; 1.05 mm) in CH₂ Cl₂ (50 ml.)was stirred at room temperature for 2 hours. It was washed with 1%NaHCO₃ solution, then with brine, dried over Na₂ SO₄ and evaporated onthe aspirator to leave 0.49 g. white solid, m.p. 146°-148° C. It wasused with no further purification. ##STR177##

A solution of isocephem compound 81 (0.49 g.; 0.9 mm) in ethyl acetate(75 ml.) was added to a prehydrogenated sample of 20% Pd(OH)₂ on carbon(0.50 g.) in ethyl acetate (25 ml.). It was then stirred under hydrogenat atmospheric pressure and after 15 minutes, gas consumption hadceased. It was filtered through a celite pad, washed well with ethylacetate, and the solvent was removed on the aspirator to leave 0.40 g.of an amorphous solid. This was suspended in ether and extracted with 2%NaHCO₃. The aqueous extract was acidified with 10% HCl and the whitesolid collected by suction filtration, washed with water and dried togive a white solid, m.p. 160°-162° with decomposition. Recrystallizedfrom CHCl₃ /ether, m.p. 162°-163° with decomposition.

Anal. Calc'd for C₂₃ H₂₂ N₂ O₆ : C, 65.39; H, 5.25; N, 6.23. Found: C,65.28; H, 5.36; N, 6.56.

A sample of compound 82 prepared above which can be named7β-phenoxyacetamido-3-phenethyl-Δ³ -O-2-isocephem-4-carboxylic acid(also called BC-L54) after solution in water and dilution with NutrientBroth was found to exhibit the following Minimum InhibitoryConcentrations (M.I.C.) in mcg./ml. versus the indicated microorganismsas determined by overnight incubation at 37° C. by tube dilution.Cephalexin was included as a comparison compound.

                  Table 4                                                         ______________________________________                                        M.I.C. in mcg./ml.                                                                                          Cepha-                                          Organism            BC-L54    lexin                                           ______________________________________                                        D. pneumoniae A9585     .03       .13                                         +5% serum*                                                                    Str. pyogenes A9604     .03       .13                                         +5% serum*                                                                    S. aureus Smith.sup.+                                                                       A9537     .06       .25                                         S. aureus Smith.sup.+                                                                       A9537     4         .5                                          +50% serum                                                                    S. aureus BX1633-2                                                                          A9606     8         1                                           at 10.sup.-3 dil'n                                                            S. aureus BX1633-2                                                                          A9606     >125      4                                           at 10.sup.-2 dil'n                                                            S. aureus meth.-                                                                            A15097    125       32                                          resist; at 10.sup.-3                                                          dil'n                                                                         Sal. enteritidis.sup.+                                                                      A9531     >125      2                                           E. coli Juhl.sup.+                                                                          A15119    .5        4                                           E. coli.sup.+ A9675     >125      8                                           K. pneumoniae.sup.+                                                                         A9977     >125      2                                           K. pneumoniae.sup.+                                                                         A15130    >125      8                                           Pr. mirabilis.sup.+                                                                         A9900     >125      4                                           Pr. morganii.sup.+                                                                          A15153    >125      >125                                        Ps. aeruginosa.sup.+                                                                        A9843A    >125      >125                                        Ser. marcescens.sup.+                                                                       A20019    >125      >125                                        Ent. cloacae  A9656     >125      >125                                        Ent. cloacae  A9657     >125      2                                           Ent. cloacae  A9659     >125      >125                                        ______________________________________                                         *50% Nutrient Broth - 45% Antibiotic Assay Broth                              .sup.+ at 10.sup.-4 dilution.                                            

EXAMPLE 7 Potassium 7β-Phenoxyacetamido-Δ³ -O-2-isocephem-4-carboxylate##STR178## Procedure

A mixture of ethyl N-formyl glycinate (21) (454 g., 3.46 moles) andethyl formate (1800 ml.) was slowly added into a suspension of freshlyprepared sodium ethoxide (3.46 moles, dried in high vacuum at 150° C.for 24 hrs; the sodium ethoxide must be completely free of ethanol asyields are considerably lowered otherwise) in 2620 ml. of dry benzenewith stirring in an ice bath for three hours. The suspension was allowedto stand at 4° C. for 18 hours. The solution was carefully decanted andthe solid residue washed several times with benzene. To the solid wasadded slowly 4500 ml. of 15% HCl-absolute ethanol. The solution wasstirred at 25° C. for 18 hours. The ethanol was removed by distillationat reduced pressure. The residue was dissolved in 4.8 liters methanol,then 750 g. of sodium bicarbonate was added at 25° C. The suspension wasstirred 18 hours, filtered and the filtrate evaporated. The residue wastaken up in 4 liters of ether, dried over anhydrous sodium sulfate andevaporated. The oily residue was distilled to give 170 g. (28%)β,β-diethoxyethylalanate (22), b.p. 90°-94° C. (0.1 - 0.13 mm).(Literature b.p. 71°/1 mm; Ellis V. Brown, Chemistry of Penicillin (H.T. Clarke et al.) Princeton Univ. Press, 1949, p. 473-534.) The NMR andIR spectra of 22 were consistent with the assigned structure.

β,β-diethoxyethylalanate was also prepared by the following procedure:

To a suspension of 49.5 g. (0.65 moles) of sodium ethoxide (Note 1) in300 ml. benzene was added a solution of 65.5 g. (0.05 moles) N-formylethyl glycinate in 300 ml. ethylformate in a 3 L. flask equipped with amechanical stirrer at 0° C. over 30 minutes. After stirrng for 1 hour,the solution clarified and was allowed to stand 18 hours at 4° C. Asolid separated from the solution. The supernatant liquid was decantedand the residue washed with 300 ml. benzene.

To the solid was added 150 ml. ethanol and 550 ml. CH₂ Cl₂ and thesuspension was cooled to 0°-5° C. in an ice bath. To the solution astream of dry HCl gas was added for 1 hour. The cooling was removed andthe HCl gas bubbled in an additional 5.5 hours after which the solutionwas allowed to stand at 25° C. for 18 hours. The excess HCl was purgedby passing a stream of nitrogen through the solution for 30 minutes andthe solution cooled to -10° C. in a methanol-ice bath. A stream ofammonia was passed through the solution until the pH = 9.0. The solutionwas diluted with 500 mg. CH₂ Cl₂ and the solid ammonium chloride removedby filtration. The filter cake was washed with an additional 500 ml. CH₂Cl₂. The filtrate was evaporated to dryness at reduced pressure (bathtemperature <45° C.). The residual oil was extracted with ether (2 × 500ml.) and the extracts evaporated to dryness. The residual oil wasextracted into petroleum ether (20-60° C.) (3 × 300 ml.). The extractswere dried over Na₂ SO₄, filtered and evaporated to yield 54.6 g.(˜54%). The NMR and IR spectra of this oil were identical in allrespects to those of authentic 22. This oil could be used as such insubsequent steps. Distillation gave 43.7 g. (42.5%) of pure 22 b.p.60°-75° C. (0.005 millimeter/Hg.). Note 1. Commercial NaOEt was used.The 30% excess was necessary. ##STR179##

Procedure A. Preparation of Schiff Base (23).

A mixture of 95.2 g. (0.46 mole) β,β-diethoxyalanine ethyl ester and60.8 g. (0.46 mole) cinnamaldehyde in 1.5 L. of methylene chloride wasboiled at reflux for 30 minutes. After this initial reflux period 850ml. of methylene chloride was distilled at atmospheric pressure over 1.5hours. (azeotrope with water). The concentrated solution was dried overanhydrous sodium sulfate (large excess >100 g.) for 1 hour. The dryingagent was removed by filtration and the solution of 23 evaporated todryness at reduced pressure and the residue pumped at <1 millimeter/Hg.pressure for 30 minutes at 40° C. The residue was then diluted to 1.3liters with dry methylene chloride.

The formation of the Schiff base may be accomplished in a number ofways. On a small scale the two reactants may be mixed in a suitablesolvent (benzene, ether, CH₂ Cl₂, etc.) in the presence of a dryingagent (Na₂ SO₄, MgSO₄). Alternatively the water may be removedazeotropically with benzene. Inasmuch as the subsequent reaction is donein methylene chloride the above method is preferred. The CH₂ Cl₂ isdried by passing thru an alumina (Act I) column which removed anyalcohol which may be present as preservative. At the end of the reactiona small aliquot was evaporated and the NMR and IR spectra taken to checkfor completeness; the yield is quantitative.

B. Preparation of azidoacetyl chloride

The azidoacetyl chloride was prepared via a modification of the methodof J. H. Boyer and J. Horner, J. Am. Chem. Soc., 77, 951 (1955).

To 128 g. (1.354 mole) chloroacetic acid in 300 ml. of water was added 7ml. 50% sodium hydroxide solution (0.0875 mole) and 110 g. (1.69 moles)of sodium azide. The slurry was contained in a three-necked 2 literround-bottomed flask fitted with two efficient condensers and anadditional funnel. [THE REACTION MUST BE DONE IN AN EFFICIENT FUME HOOD!The original literature preparation used at least one equivalent ofsodium hydroxide whereas we used less than 10 mole %.] The mixture waslayered with 100 ml. ether and heated on a steambath for 24 hours. Theorange solution (occasionally colorless) was cooled to 0°-5° C. in anice bath. To the cooled solution was added 300 ml. of 10% H₂ SO₄followed by solid sodium chloride to saturation. The solution wasextracted with ether (5 × 200 ml.), the extracts dried over sodiumsulfate. The drying agent filtered, and the ether evaporated below 30°C. at reduced pressure. The NMR spectrum of the residual oil indicatedit to be a 1/2 hydrate of azidoacetic acid contaminated with 10% ether.The oil was used without further purification.

To the oil cooled in an ice bath was added 340 g. (2.85 moles) thionylchloride (the addition of the first 50 ml. is slow as vigorous gasevolution occurs, the remainder is added quickly). After addition, thesolution was refluxed 2 hours. The excess thionyl chloride was distilledat reduced pressure (<50° C., 70 mm). The residue was distilled to yield93 g. (57.2%) azidoacetyl chloride, b.p. 38°-40° C/12-15 mm Hg. Thedistillation must be carried out using a water bath the temperature ofwhich must never exceed 80° C. An explosion occurred in one run wherehigher bath temperatures were utilized. The NMR and IR spectra areconsistent with the assigned structure.

C. β-lactam formation

The freshly prepared Schiff base solution (0.46 moles in 1.3 l.methylene chloride) was cooled to 0°-5° C. with ice bath. To this 46.1g. (0.46 mole) triethylamine was added. A solution of 56.0 g. (0.46moles) azidoacetyl chloride in 500 ml. methylene chloride was addeddropwise over 1 hour. The solution was stirred an additional 30 min.,washed with water, saturated NaCl solution, and dried over Na₂ SO₄.Evaporation of this solution yielded 187.0 g. (>98% crude yield) ofcompound 24 as a reddish oil.

The compound 24 was obtained as a mixture of diastereoisomers (24a and24b). Occasionally a by-product was also obtained when excesses of azidoacetyl chloride were used. This by-product has been identified as 25.##STR180##

Compound 25 is thought to arise from further reaction of 24a and/or 24bwith the acid chloride. The stereochemistry of the azido and styrrylsubstitutents of 24a, 24b and 25 has been shown to be exclusively cis;no trace of trans component could be detected by NMR. The mixture ofcompounds 24a and 24b was readily separated from 25 by columnchromatography (Dry-column technique on silica gel (15% H₂ O) usingether as eluent).

Purification of compounds 24a and 24b by chromatography was notsatisfactory as losses occurred. Only partial separation could beachieved. When the cycloaddition was carried out as described formationof 25 was minimal (<2 - 5%).

Compound 25 was shown to be a single geometrical isomer as indicated inthe diagram above.

Generally the compound 24 was used as such in subsequent reactions.

A small sample of the crude β-lactam 24 was chromatographed on silicagel (deactivated -- 15% water) by dry column technique using methylenechloride as eluent. Two pure fractions (as determined by TLC and NMR)were obtained corresponding to compounds 24 and 25. The oils wereanalyzed. Compounds 24 (mixture of diastereoisomers) MW = 402.460

Anal. Calc'd for C₂₀ H₂₆ N₄ O₅ . 0.1 CH₂ Cl₂ : C, 58.50; H, 6.35; N,13.64. Found: C, 58.48; H, 6.48; N, 13.38. Compound 25 MW = 356 390

Anal. Calc'd for C₁₈ H₂₀ N₄ O₄ : C, 60.66; H, 5.66; N, 15.72. Found : C,60.78; H, 5.73; N, 15.91. ##STR181##

Procedure

A. In a 3 liter round bottomed flask equipped with a magnetic stirrerwas dissolved 119 g. (0.296 moles) of compound 24 in 1 liter of ethanol.To this was added 100 ml. of water followed by 1230 ml. of 1% aqueoussodium hydroxide over a period of 1.5 hours. The solution was maintainedat 20°-25° C. After addition, the solution was stirred 1 hour. To thesolution was added 10% HCl to pH 3 followed by 500 ml. saturated NaClsolution. The aqueous phase was extracted by three portions of CHCl₃(600, 300, 300 ml.) and the organic layer evaporated. The residue wasre-dissolved in 500 ml. ether and extracted with three portions (600,300, 300 ml.) of saturated sodium bicarbonate solution. The aqueouslayer was acidified to pH 3 with 10% HCl and extracted into CHCl₃ (inthree portions, 600, 300 and 300 ml.). The organic layer was dried overMgSO₄, filtered, and evaporated to yield 97 g. (88% yield) of crude acid26. The IR and NMR spectra of the acid were consistent with the assignedstructure.

B. The acid 26 (192 g.) was dissolved in 1 l. dry CH₂ Cl₂ with 53.5 g.(73 ml.) of triethylamine and cooled to 0°-5° C. in an ice bath. To thiswas added benzyl chloroformate (96 g.) dropwise over a two hour periodwith stirring. Following the addition the solution was stirred at roomtemperature for 30 min. The solution was washed with water (2 × 200 ml.)until neutral, with brine solution (100 ml.) and then dried over MgSO₄.Evaporation afforded 27 as a dark brown oil. The oil was passed througha column of granular adsorbant magnesium silicate (Florisil; 400 g.)with methylene chloride to give 204 g. (85% crude yield) of the desiredester 27. NMR and IR spectra were compatible with the assignedstructure.

Generally the oil was not purified further but was used as such insubsequent reactions.

The reaction proceeds according to the following scheme. ##STR182##

The IR spectrrum of the crude reaction mixture indicates no mixedanhydride to be present.

The crude product contained the mixture of desired diastereoisomericesters, benzyl alcohol, and a small amount of acid. The columnchromatography removed much of the acid and other impurities. The benzylesters are generally not very stable to column chromatography sopurification is usually carried out at a later step in the sequence.##STR183##

Procedure

A solution of 4.8 g. (10.04 mmoles) compound 27 in 80 ml. dry methylenechloride was prepared and cooled to -78° C. on an acetone-dry ice bath.To this was added ozone until a blue color persisted. The ozone additionwas ended and the excess ozone removed by bubbling dry nitrogen throughthe solution. To the solution was added 5 ml. of dimethyl sulfide.

The purpose of the dimethyl sulfide is to decompose the initially formedozonide. ##STR184## The DMSO thus produced can also react with theozonide as shown below ##STR185## The washing with the NaHCO₃ removesthe benzoic acid thus produced.

The solution was allowed to come to room temperature over 1 hour. Thesolution was then washed with water (20 ml.), saturated NaHCO₃ (20 ml.),water (10 ml.), brine, and dried over MgSO₄. The solution was filteredand evaporated to give 5.0 g. of an oil. The by-product benzaldehyde wasremoved by distillation at 0.05 mm. Hg. and a bath temperature of ˜65°C. The residual oil 4.0 g (95%) was analyzed by NMR which indicates 77%free aldehyde 28.

The desired aldehyde forms a hydrate which tends to lower the amount offree aldehyde observable in the NMR spectrum. ##STR186##

Procedure

To 3.5 g. (9.0 mmoles) of compound 28 in 30 ml. 95% ethanol at 0°-5° C.was added 255 mg. (6.0 mmoles) of sodium borohydride with stirring.After 30 min. at 0°-5° C. the solution was stirred an additional 30 min.at 25° C. The solution was acidified to about pH 4 with 10% hydrochloricacid and diluted with 40 ml. ice water. The aqueous layer was extractedwith chloroform (3 × 30 ml.). The combined extracts were washed withwater (2 × 10 ml.), brine, dried over MgSO₄, filtered and evaporated toyield 3.4 g. crude alcohol 29. The oil was chromatographed on silica gel(5% water) with chloroform to yield 3.0 g. pure alcohol 29 (85%). The IRand NMR spectra were compatible with the assigned structure. ##STR187##

Procedure

A mixture of 3.2 g. (8.17 mmoles) compound 29, 11 ml. acetic anhydride,and 1.12 g. (8.2 mmoles) zinc chloride was stirred 18 hours at 25° C.The reaction mixture was evaporated at reduced pressure and the residuetaken up in 50 ml. -- methylene chloride -- 20 ml. water. The organicphase was separated, washed with water, brine, dried over MgSO₄,filtered and the filtrate evaporated to yeild 3.0 g. of an oil. The oilwas chromatographed on 50 g. silica gel (deactivated -- 5% water) by drycolumn technique using chloroform as an eluent. Evaporation of theeluent gave 1.3 g. (41%) of pure 30 as an oil. The IR and NMR spectrawere compatible with the assigned structure. ##STR188##

Procedure

Compound 30, 5.95 g. (15.35 mmoles) was refluxed in 35 ml. CH₃ OH and 35ml. 10% hydrochloric acid for a period of 1 hour. The solvent waspartially evaporated at reduced pressure and the aqueous residue wasextracted with chloroform (3 × 30 ml.). The combined extracts werewashed with water (2 × 10 ml.), saturated brine, and dried overanhydrous magnesium sulfate. The solution was filtered and evaporated togive 4.6 g. (87% yield) of 31 as an oil. The NMR and IR spectra of thisoil were consistent with the assigned structure. ##STR189##

Procedure

A solution of 4.6 g. (13.3 mmoles) compound 31, 1.03 g. (14.0 mmoles)pyrrolidine, and 900 mg. (14.0 mmoles) acetic acid in 50 ml. of benzenewas refluxed 18 hours. The solvent was evaporated at reduced pressureand the residual oil was taken up in 60 ml. of chloroform. Thechloroform solution was washed with water (15 ml.), brine, and driedover MgSO₄. The drying agent was filtered off and the filtrateevaporated to dryness to give 3.50 g. (71% yield) of crystalline enamine32, m.p. 111.5° - 112.5° C. The NMR and IR spectra were compatible withthe assigned structure.

Anal. Calcd for C₁₈ H₂₁ N₅ O₄ : C, 58.21; H, 5.70; N, 18.86. Found: C,58.23; H, 5.72; N, 19.10. ##STR190##

Procedure

A solution of 2.44 g. (6.6 mmoles) compound 32, 3.92 g. (33 mmoles)methane sulfonyl chloride and 3.3 g. (33 mmoles) triethylamine in 50 ml.methylene chloride was stirred at ambient (25° C.) temperature for 74hours. The reaction mixture was washed with water (2 × 10 ml.), brine,and dried over Na₂ SO₄. The drying agent was filtered off and thefiltrate evaporated to dryness. The oil was filtered through a silicagel column (deactivated - 15% water) (16 g.) with chloroform to give 2.6g. (90%) of crystalline mesylate 33, m.p. 116° - 117.5° C. The IR andNMR spectra were compatible with the assigned structures. ##STR191##

Procedure

A solution of 2.28 g. (5.26 mmoles) compound 33 in 25 ml. of acetone and25 ml. 10% hydrochloric acid was refluxed 15 minutes. The acetone wasevaporated at reduced pressure and the residue extracted with chloroform(3 × 30 ml.). The chloroform layer was washed with water and evaporatedto dryness. The residual oil was dissolved in ether (20 ml.) and thesolution extracted with saturated sodium bicarbonate solution (4 × 8ml.). The bicarbonate was acidified to pH 4 with 10% HCl andre-extracted with chloroform (3 × 50 ml.). The chloroform was washedwith water, brine and dried over MgSO₄. The drying agent was filteredoff and the filtrate evaporated to give 1.62 g. (81%) of compound 34.The IR and NMR spectra of 34 were compatible with the assignedstructure.

The NMR spectrum of 34 shows two signals for the benzyl group. This maybe due to two causes - hydrogen bonding causing restricted rotation orgeometrical isomerism. ##STR192##

Procedure

To a suspension of 198 mg (4.70 mmoles) sodium hydride (55% mineral oildispersion, washed 3X with petroleum ether) in 5 ml. dry dimethylsulfoxide (DMSO) was added a solution of 1.62 g. (4.27 mmoles) compound34 in 5 ml. DMSO over 5 min. with stirring at 25° C. [Gas evolution wasobserved to cease after 15 - 20 minutes. Prolonged reaction times gavelower yields of 35. The optimum time was 45-60 minutes.] After 1 hour,the reaction mixture was poured into 50 ml. 1% HCl-ice water and wasextracted with chloroform (4 × 30 ml.). The organic layer was washedwith water (3 × 10 ml.) brine, and dried over MgSO₄. Filtration andevaporation of the filtrate gave 1.2 g. of 35 as an oil. Triturationwith ether caused crystallization; 545 mg., m.p. 110°, of 35 werecollected. The NMR and IR spectra were compatible with the assignedstructure.

Anal. Calcd. for C₁₄ H₁₂ N₄ O₄ : C, 55.99; H, 4.03; N, 18.66. Found: C,55.23; H, 4.02; N, 18.91. ##STR193##

Procedure

A. Compound 35 (500 mg.; 1.66 mmoles) was dissolved in 20 ml. of dryethyl acetate. To this was added 450 mg. of 10% Pd/C and the solutionwas stirred under hydrogen at atmospheric pressure and room temperaturefor 30 min. The solution was filtered through diatomaceous earth("Celite") and the filter cake washed thoroughly with methyllenechloride. Evaporation of the filtrate yielded 500 mg. of crude amine 36.The NMR and IR spectra of the compound were compatible with the assignedstructure. Compound 36 was used in the subsequent step without furtherpurification. [On standing some decomposition was noted. The amineshould be used as soon as possible after preparation.]

B. Compound 36 (500 mg.) was dissolved in 10 ml. of dry methylenechloride and cooled to 0°-5° C. in an ice bath. To this was added 280mg. (2.8 mmoles) of triethylamine and 346 mg. (2.0 mmoles) ofphenoxyacetyl chloride was added slowly. After stirring for 1 hour at0°-5° C. the solution was washed with water (2 × 10 ml.) and dried overNa₂ SO₄. After evaporation the residual oil was taken up in 50 ml. ofether and filtered. The filtrate was evaporated and triturated withether-petroleum ether (1:1). The solid thus obtained was collected byfiltration to yield 570 mg. crude amide 37. The amide waschromatographed on a silica gel column (undeactivated) (25 g.) withbenzene-acetone (initially in a ratio 50:1, gradually changed to 1:1 -2% more acetone every 25 ml.). The desired amide 37 was obtained pure,195 mg.

The NMR and IR spectra were compatible with the assigned structure.##STR194##

Procedure

Compound 37 (210 mg.; 0.514 mmoles) was dissolved in 40 ml. ethylacetate and 1 ml. glacial acetic acid was added. Using 610 mg. (˜ 20%)palladium hydroxide on charcoal as catalyst, the solution washydrogenated at 58 psi for 50 minutes.

The reaction mixture was filtered through "Celite" (twice) and thecatalyst was washed thoroughly with chloroform (20 ml.). The filtratewas evaporated to dryness and then evaporated 3 times with benzene inorder to strip off the acetic acid. A very viscous oil was obtainedwhich was washed with 10 ml. benzene. The residual oil was scratchedwith 10 ml. ether. The solid 38 which formed was collected by filtrationto yield 115 mg. (70.5%). U.V. λmax. 268, ε = 9549. No sharp m.p. wasobserved. (D.p. <252° C.)

Anal. Calcd. for C₁₅ H₁₄ N₂ O₆. 1/2 hydrate: C, 55.06; H, 4.62; N, 8.56.Found: C, 55.19; H, 4.70; N, 9.00. ##STR195##

Procedure

To a solution of 30 mg. compound 38 in 3 ml. methyl isobutyl ketone wasadded one or two drops of 50% solution of potassium 2-ethylhexanoate inbutanol. A white crystalline material separated almost immediately whichwas collected by filtration, washed with methyl isobutyl ketone anddried over P₂ O₅ for 48 hours under high vacuum to yield 18 mg. 39(53.5%). U.V. λmax. 263, ε 5528. No sharp m.p. or d.p. could beobserved.

Anal. Calcd. for C₁₅ H₁₃ N₂ O₆ K.1/2 H₂ O: C, 49.31; H, 3.82; N, 7.67.Found: C, 49.35; H, 3.94; N, 8.21.

A sample of compound 39 prepared above which can be named potassium 7β-phenoxyacetamido-Δ³ -O-2-isocephem-4-carboxylate (called BC-L2) aftersolution in water and dilution with Nitrient Broth was found to exhibitthe following Minimum Inhibitory Concentrations (M.I.C.) in mcg./ml.versus the indicated microorganisms as determined by overnightincubation at 37° C. by Tube Dilution. One old, orally absorbedcephalosporin (cephalexin) was included.

                  Table 5                                                         ______________________________________                                        M.I.C. in meg./ml.                                                                                          Cepha-                                          Organism            BC-L2     lexin                                           ______________________________________                                        D. pneumoniae A9585     .6        .6                                          +5% serum*                                                                    Str. Pyogenes A9604     .6        .3                                          +5% serum*                                                                    S. aureus Smith+                                                                            A9537     1.3       1.3                                         S. aureus Smith                                                                             A9537     1         2.5                                         +50% serum                                                                    S. aureus BX1633-2                                                                          A9606     2.5       4                                           at 10-3 dil'n                                                                 S. aureus BX1633--2                                                                         A9606     >125      8                                           at 10.sup.-2 dil'n                                                            Sal enteritidis                                                                             A9531     16        4                                           F. coli Juhl  A15119    63        8                                           F coli        A9675     >125      16                                          K. pneumoniae+                                                                              A9977     32        8                                           D. pneumoniae+                                                                              A15130    >125      16                                          Pr. mirabilis A9900     63        4                                           Pr. morganii+ A15153    >125      >125                                        Ps. aeruginosa+                                                                             A9843A    >125      >125                                        Ser. marcescens+                                                                            A20019    >125      >125                                        S. aureus meth.                                                                             A15097    >125      32                                          resist; at 10.sup.-3                                                          dil'n                                                                         ______________________________________                                         *50% Nutrient Broth - 45% Antibiotic Assay Broth                              + at 10.sup.-4 dilution                                                  

EXAMPLE 8 ##STR196## Procedure

A solution of 43.0 g. (0.107 mole) of compound 24 (as prepared by themethod of Example 7) in 700 ml. of dry methylene chloride was cooled to-78° C. in an acetone dry ice bath and a steam of ozone passed throughfor 2 hours. At the end of this time the solution turned bluish-greenand the ozone was replaced by a stream of dry nitrogen. When the excessozone had been purged (as indicated by the disappearance of the bluecolor) 30 ml. of dimethyl sulfide was added. The solution was allowed tocome to room temperature (˜ 25° C.) over 1 hour. The solution wasevaporated to dryness and the residue redissolved in 800 ml. CH₂ Cl₂.The solution was washed with water, brine and dried over MgSO₄.Evaporation of the solution gave an oily residue which was distilled for18 hours at 40° - 50° C. and 0.1 mmHg to remove benzaldehyde. Thisyielded 40.5 g. of 40 as an oil. The NMR spectrum indicated 76% freealdehyde. The crude aldehyde was used in the next step without furtherpurification. ##STR197##

Procedure

A solution of 40.3 g. of compound 40 in 250 ml. ethanol -12.5 ml. H₂ Owas prepared and cooled to 0°-5° C. in an ice bath. To this was added1.56 g. (0.041 moles) sodium borohydride and the solution was stirredfor 30 min. at 0°-5° C. To the solution was added 10% hydrochloric acidto pH 4. The reaction mixture was evaporated to dryness at reducedpressure below 35° C. To the residue was added 200 ml. of brine and thesolution was extracted with chloroform (3 × 200 ml.). The extracts weredried over Na₂ SO₄, filtered and evaporated to yield 37.0 g. of crudealcohol 41. The crude alcohol was filtered through a column of activityIII alumina (550 g.) using chloroform as an eluent to yield 27.0 g. ofreasonably pure alcohol 41 (>90%). The IR and NMR spectra of the oilwere compatible with the assigned structure.

The overall yield from compound 24 was 76.5%. It has been found that theozonolysis and reduction procedure can be combined by carrying out theoxidation in ethanol and reducing the ozonide in situ with NaBH₄. Thisgives an 83-85% yield of equimolar amounts of 41 and benzyl alcohol. Asmall sample of the alcohol was purified by column chromatography onalumina (Act II). MW - 330.351.

Anal. Calcd. for C₁₃ H₂₂ N₄ O₆ : C, 47.27; H, 6.71; N, 16.96. Found: C,47.26; H, 6.85; N, 17.15. ##STR198##

Procedure

To a solution of 6.20 g. (18.8 mmole) of alcohol 41 in 100 ml. of drymethylene chloride was slowly added a solution of 4.0 g. (28.2 mmole,3.54 ml.) boron trifluoride etherate in 20 ml. dry methylene chlorideover 15 min. at 0°-5° C. The cooling bath was removed and stirring wascontinued for 18 hours. The reaction mixture was filtered through acolumn of activity III alumina (40 g.). The column was washed with 300ml. chloroform. The eluted fractions were evaporated to dryness to yield6.0 g. of 42 as an oil which by TLC analysis was at least 90% pure. TheNMR and IR spectra were compatible with the assigned structure [Incontrast to the methyl ester dimethyl acetal analog of 41 only oneisomer was obtained on cyclization. Careful chromatography of 42 gave a76% yield of one pure isomer with the stereochemistry indicated in thefigure. A small sample was rechromatographed for analysis.

Anal. Calcd for C₁₁ H₁₆ N₄ O₅ : C, 46.47; H, 5.67; N, 19.71. Found: C,46.54; H, 5.85; N, 19.34. ##STR199##

Procedure

To a solution of 12.2 g. (43 mmoles) of compound 42 in 180 ml. ethanolwas added 175 ml. 1% sodium hydroxide over a period of 10 min. at <25°C. The solution was stirred an additional 20 minutes. The ethanol wasevaporated at reduced pressure and the alkaline solution was extractedwith ether (2 × 200 ml.). The organic layer was discarded and theaqueous solution acidified to pH 3-4 with 10% hydrochloric acid. Thesolution was extracted with chloroform (2 × 100 ml.), the organic layerwashed with water (50 ml.), brine (50 ml.), and dried over MgSO₄.Evaporation gave 7.25 g. (66%) of acid 43. Trituration with ether andfiltration gave pure acid 43, m.p. 114°-115° C. The NMR and IR spectrawere compatible with the assigned structure.

Anal. Calcd. for C₉ H₁₂ N₄ O₅ : C, 42.19; H, 4.72; N, 21.87. Found: C,42.18; H, 4.83; N, 22.01. ##STR200##

Procedure

A mixture of 760 mg. (2.8 mmole) of compound 42, 925 mg. ammoniumchloride (17.1 mmoles) and 620 mg. (17.1 mm.) zinc powder in 35 ml.ethanol was stirred at 25° C. for 3 hours. The reaction mixture wasfiltered through "Celite" and the filtrate evaporated to dryness. Theresidue was taken up in chloroform and filtered through 30 g. of Alumina(Act III). Evaporation of the eluent yielded 578 mg. of crude amine 44.The amine was redissolved in 15 ml. chloroform and extracted into 10%HCl (2 × 3 ml.). The aqueous layer was neutralized with sodiumbicarbonate and extracted into chloroform. The extracts were dried overNa₂ SO₄, filtered, and extracted into chloroform. The extracts weredried over Na₂ SO₄, filtered, and evaporated to yield 360 mg. of an oilwhich crystallized on standing. The amine 44 was recrystallized fromether, m.p. 98.5° - 99° C. The IR and NMR spectra were compatible withthe assigned structure. MW = 258.

Anal. Calcd. for C₁₁ H₁₈ N₂ O₅ : C, 51.15; H, 7.03; N, 10.85. Found: C,51.16; H, 7.01; N, 11.03. ##STR201##

Procedure

A solution of 400 mg. (1.55 mmole) of compound 44, 410 mg. (1.64 mmole)EEDQ and 250 (1.69 mmole) phenoxyacetic acid in 20 ml. dry methylenechloride was stirred at 25° C. for a period of 1.5 hours. The reactionmixture was filtered through a column of alumina (activity III 8 g.) andthe eluent evaporated to dryness. The result solid was washed with etherand collected by filtration to yield 554 mg. (90%), m.p. 162°-164°.Recrystallization from chloroform-ether gave analytically pure amide 45,m.p. 166.5°-167.5° C. The IR and NMR spectra were compatible with theassigned structure. NW = 392.417

Anal Calcd. for C₁₉ H₂₄ N₂ O₇ : C, 58.15; H, 6.16; N, 7.14. Found: C,57.93; H, 6.23; N, 7.34. ##STR202##

Precedure

A solution of 392 mg. (1 mmole) compound 45 in 13 ml. warm methanol wasassed to 12 ml. 1% NaOH at 25° C. with stirring over 10 min. After 1hour the methanol was evaporated and the alkaline solution was extractedwith chloroform (2 × 20 ml.). The aqueous solution was acidified to pH ˜4 with 10% HCl and extracted with chloroform (2 × 15 ml.). The extractswere washed with water, dried over Na₂ SO₄, filtered, and evaported togive 200 mg. (55%) of a white solid. Recrystallization frommethanol-ether gave pure acid 46, m.p. 150°-151.5°0 C. The IR and NMRspectra were compatible with the assigned structures. MW 32 364.363

Anal. Calcd. for C₁₇ H₂₀ N₂ O₇.0.5 CH₃ OH: C, 55.26, H, 5.83; N, 7.37.Found : C, 55.08; N, 5.53; N, 7.48. ##STR203##

Procedure

To a solution of 6.25 g. (25.6 mmoles) of compound 43 in 100 ml. ofether was added 5.35 g. (25.6 mmoles) phosphorous pentachloride. Thesuspension was refluxed for 15 min. after which the clear solution wasdecanted and evaporated to dryness. The residual oil was taken up in 50ml. benzene and evaporated to dryness at reduced pressure. Thisprocedure was repeated three times to remove phosphorous oxychloride.The residual oil was then pumped in high vacuum (0.05 mm Hg) at 30° C.for 1 hour. The NMR and IR spectra were compatible for the desired acidchloride.

The acid chloride was taken up in 20 ml. dry methylene chloride and wasadded to a mixture of 2.7 g. (26 mmole) benzyl alcohol and 3.2 g. tri -ethylamine in 50 ml. dry methylene chloride at 25° C. over a period of10 minutes. The solution was stirred for 1 hour, washed with water (2 ×20 ml.), brine and filtered through 20 g. of "Florisil". The eluent wastreated with charcoal (Norite), dried over MgSO₄, filtered andevaporated to give 7.4 g. (83.5%) of crude benzyl ester 47. Triturationwith benzenepetroleum ether caused crystallization. The solid wasrecrystallized from benzene-petroleum ether to yield pure 47, m.p.79°-79.5° C. The IR and NMR spectra were compatible with the assignedstructure. MW = 346.352

Anal. Calcd. for C₁₆ H₁₈ N₄ O₅ : C, 55.49; H, 5.24; N, 16.18. Found: C,55.81; H, 5.36; N, 16.40. ##STR204##

Procedure

A mixture of 6.5 g. (18.8 mmole) compound 47, 50 ml. acetic anhydride,and 5.1 g. (37.6 mm) zinc chloride was stirred at 0°-5° C. for 30 min.then at 25° C. for 18 hours. The reaction mixture was evaporated todryness at reduced pressure and the residue taken up in 200 ml.methylene chloride-50 ml. water. The organic phase was separated, washedwith water, brine, and dried over MgSO₄. Filtration and evaporation ofthe filtrate gave an oily residue which was chromatographed on 60 g.alumina (Activity III) with chloroform to yield 5.35 g. (73%) ofcompound 30 identical in all respects with that obtained earlier.##STR205##

Procedure

To 325 mg. (0.84 mmole) of compound 30 in 5 ml. dry methanol was added325 mg. zinc powder and 300 mg. ammonium chloride at 0°-5° C. Thesuspension was stirred for 1 hour, filtered, and the filtrate evaporatedto yield 312 crude amine 48. The IR spectrum indicated completereduction of the azido function.

The crude amine 48 was treated with 140 mg. (0.92 mmole) phenoxyaceticacid and 230 mg. (0.92 mmole) EEDQ in 10 ml. methylene chloride at 25°C. for 1 hour. The solution was washed with 10% HCl (5 ml.), water (5ml.), brine, dried over Na₂ SO₄, filtered, and the filtrate evaporated.The crude amide 49 was chromatographed on alumina (Activity III) usingchloroform as eluent to give 230 mg. pure amide 49 and an additional 100mg. of ˜80% pure amide. The NMR and IR spectra were compatible with theassigned structures. ##STR206##

Procedure

A solution of 110 mg. (0.222 mmole) of 49 in 4 ml. methanol and 2 ml.10% hydrochloric acid was refluxed for 1 hour, diluted to 20 ml withwater and extracted into chloroform. The extracts were dried over Na₂SO₄, filtered, and evaporated to yield 74 mg. (75%) of the desiredalcohol 50. The NMR and IR spectra were compatible with the assignedstructure. ##STR207##

Procedure

A solution of 140 mg. (0.31 mmole) compound 50, 140 mg. acetic acid, and140 mg. pyrrolidine in 10 ml. benzene was boiled are reflux for 16hours. The solution was evaporated to dryness, taken up in chloroform(20 ml.), washed with water (5 ml.), 10% HCl (5 ml.), saturated NaHCO₃solution (5 ml.), brine (5 ml.), dried over Na₂ SO₄, filtered, andevaporated to yield 157 mg. crude enamide 51 (100%). The IR and NMRspectra were compatible with the assigned structure. ##STR208##

Procedure

To a solution of 157 mg. (0.31 mmole) of crude 51 and 0.5 ml. pyridinein 5 ml. methylene chloride was added 170 mg. of methane sulfonylchloride at 0°-5° C. The solution was stirred at 25° C. for 48 hours,evaporated at reduced pressure, taken up in chloroform, washed withwater and dried over Na₂ SO₄. The solution was filtered and passedthrough a short column of alumina (5 g. Activity III) with chloroform aseluent. Evaporation of the eluted fraction gave 170 mg. (>90%) crudemesylate 52. The NMR and IR spectra of 52 were compatible with theassigned structure. ##STR209##

Procedure

A solution of 200 mg. compound 52 in 6 ml. acetone and 1.5 ml. 10% HClwas refluxed for 15 min., diluted to 50 ml. with water and extractedinto chloroform (3 × 25 ml.). The extracts were evaporated and theresidue redissolved into chloroform which was extracted with sodiumbicarbonate solution. The aqueous layer was acidified to pH ˜ 4 with 10%HCl and extracted into chloroform (3 × 25 ml.). The extracts were driedover Na₂ SO₄, filtered, and evaporated to yield 180 mg. crude enol 53.The NMR and IR spectra were compatible with the assigned structure.##STR210##

Procedure

Compound 53 (70 mg.) was treated for 1 hour with 5.6 mg. (60% mineraloil dispersion - washed with petroleum-ether) of sodium hydride in 2 ml.DMSO at 25° C. The solution was poured into 20 ml. ice cold 1% HCl andextracted into chloroform (3 × 10 ml.). The solution was washed withwater (2 × 10 ml.), brine (10 ml.), dried over Na₂ SO₄, filtered, andevaporated to give 60 mg. of crude 37 identical in all respects to thatobtained via acylation of 36.

EXAMPLE 9 Benzyl 7β-Azido-Δ³ -O-2-isocephem-4-carboxylate ##STR211##

A solution of 682 mg. (0.00174 moles) of 34 and 195 mg. (0.00192 moles)TEA in 10 ml. CH₂ Cl₂ was refluxed for 5 hours. The solution was washedwith water, brine, dried over Na₂ SO₄, filtered and evaporated to yieldan oil. Trituration with ether and filtration gave 393 mg. (75.5%) pure35. An additional 21.0 mg. were recovered from the mother liquors.Compound 35 prepared in this manner was identical in all respects withthat obtained earlier (m.p., IR and NMR).

EXAMPLE 10 Ethyl 7β-Azido-Δ³ -O-2-isocephem-4-carboxylate ##STR212##

To a solution of 9.6 g. (29 mmole) of compound 83 (note 1) and 9.0 ml.of triethylamine in 96 ml. of methylene chloride at 0°-5° was addeddropwise a solution of 4.8 g. (42 mmole) of methanesulfonyl chloride in24 ml. of methylene chloride. After standing at 25° for 1 hour, thesolution was washed with equal volumes of water and 10% hydrochloricacid. Evaporation of the solvent gave a yellow oil which waschromatographed on 180 g. of alumina (grade III). Elution withchloroform gave the partially purified product 84 as a yellow oil, 5.56g. (47% yield). The IR and the NMR were consistent with the assignedstructure. (Note 2)

Note 1: The purity of the starting material (compound 83) was not knownwith certainty but it may have been less than 70% pure.

Note 2: The NMR indicated the product was of about 70% purity.##STR213##

A mixture of 4.9 g. (12 mmole) of compound 84, 10 ml. of aceticanhydride, 10 ml. of acetic acid and 1.75 g. (13 mmole) of zinc chloridewas stirred at 25° for 17 hours, then evaporated to a tar. A methylenechloride solution of the tar (50 ml.) was washed with equal volumes ofwater, 5% sodium bicarbonate and dilute sodium chloride. The methylenechloride solution was filtered through 15 g. of alumina (grade III) andevaporated to give an oil. Trituration of the oil with ether gave purecompound 85 as a colorless powder, 1.88 g. (45% yield). The IR and NMRwere consistent with the assigned structure. ##STR214##

To a solution of 1.83 g. (5.99 mmole) of compound 85 in 20 ml. oftetrahydrofuran was added 20.0 ml. of 0.25M sodium hydroxide (Note 1)solution dropwise over 10 minutes. The resulting solution wasconcentrated to 20 ml. on the rotary evaporator at 30°. The concentratewas washed with chloroform (3 × 10 ml.) (Note 2). The aqueous layer wasevaporated to dryness under high vacuum. The resulting residue (sodiumenolate 86 ) was stirred with 7.5 ml. of dimethyl sulfoxide for onehour. Water (30 ml.) and saturated sodium chloride (40 ml.) followed bya few drops of 10% hydrochloric acid were added to the dimethylsulfoxidesolution. The resulting mixture was extracted with chloroform (3 × 40ml.) and the combined chloroform layers were washed with water andevaporated to give the crude product. Pure compound 87 was obtained byrecrystallization from benzene/cyclohexane, then chloroform, ascolorless crystals, 0.39 g. (33% yield). The IR and NMR were consistentwith the assigned structure.

Note 1: Other concentrations of base and other solvents (acetone,dimethoxyethane, acetonitrile) were tried but the conditions describedhere gave better yields.

Note 2: The chloroform extract gave a yellow oil, 0.58 g. containing 65%compound 85 and 35% of an unidentified byproduct.

EXAMPLE 11 Benzyl 7β-Azido-Δ³ -O-2-isocephem-4-carboxylate ##STR215##

To a solution of 260 mg. (0.64 mmole) of compound 88 in 2.5 ml. oftetrahydrofuran was added 2.55 ml. of 0.25 M sodium hydroxide solutiondropwise over 10 minutes. The solution was concentrated to 2 ml. on therotary evaporator. The concentrate was washed with chloroform (2 × 2ml.), then evaporated to dryness under high vacuum. The residue wasstirred with one ml. of dimethyl sulfoxide for one hour. Water (1 ml.),saturated sodium chloride (1 ml.) and one drop of 10% hydrochloric acidwere added. The mixture was extracted with chloroform (3 × 2 ml.) andthe combined chloroform layers were washed with water and evaporated togive crude compound 89 as a yellow solid, 103 mg. (54% yield). The IRand NMR were consistent with the assigned structure. The NMR indicatedthe product was about 75% pure (i.e. true yield of 40%).

EXAMPLE 12 7β-Phenoxyacetamido-3-carbomethoxymethylene-Δ³-O-2-isocephem-4-carboxylic acid ##STR216##

Apparatus consisting of a 250 ml. three necked flask equipped with a lowtemperature therometer, a gas inlet, protected with a gas bubbler(paraffin oil) and magnetic stirring, was dried by heating with a bunsentorch while passing dry nitrogen through the apparatus. It was allowedto cool to 25° C. before being opened, under nitrogen flow, to introducethe reagents.

A solution of benzy 7-β-([aminophenoxyacetoyl]-3-methyl-Δ³-O-2-isocephem-4-carboxylate 90 (2.11 g.; 5 mmole) in 100 ml. of THF¹was cooled to -70° under a slow nitrogen stream. A solution of 1.66 Mbutyl lithium² (6.34 ml.; 10.5 mmole) was slowly added keeping thereaction temperature at -70° C. A slow stream of dry carbon dioxide gaswas then introduced into the reaction mixture, the cooling bath wasremoved, and the carbon dioxide introduction continued until thereaction temperature reached 25°.

The reaction mixture was poured into 200 ml. of 10% hydrochloric acid,saturated with sodium chloride, and extracted three times withdiethylether (150 ml. portions). The conbined extracts were washed threetimes with brine, dried (anhydrous sodium sulfate) and evaporated invacuo to give 2.09 g. of a yellow gum. This gum was partitioned betweendiethylether and 10% sodium bicarbonate solution three times. Thebicarbonate solutions were then washed with diethylether (twice; 50 ml.portions) and with methylene chloride (twice; 50 ml. portions). Thebicarbonate solution was then acidified to pH2 with concentratedhydrochloric acid and extracted three times with methylene chloride (100ml. portions). The methylene chloride extracts were washed twice withbrine, dried (anhydrous sodium sulfate), and evaporated in vacuo to give0.31 g. of a colorless gum. This gum was used as such in the next step.##STR217##

A solution of diazomethane in diethylether¹ was slowly added to asolution of benzyl 7-B-[aminophenoxyacetoyl]-3-carboxymethylene-Δ³-O-2-isocephem-4-carboxylate 91 (0.88 g.) in 100 ml. of diethylether,until a permanent yellow color (excess diazomethane) was produced. Thereaction mixture was then stirred at room temperature for 10 minutes.The reaction was acidified with 10% hydrochloric acid and extractedtwice with 100 ml. portions of diethylether. The extract was washed with10% sodium bicarbonate solution (twice; 75 ml. portions), with brine(once; 100 ml.), dried (anhydrous sodium sulfate), and evaporated invacuo to give 0.72 g. of crude product and then dry columnchromatographed over 36 g. of activity III silica gel. Elution withchloroform gave a fraction containing 240 mg. of methyl ester 92 NMR andIR spectra are in agreement with the assigned structure. This materialwas used as such in the next step.

A mixture of benzyl 7-β-[aminophenoxyacetoyl]-3-carbomethylene-Δ³-O-2-isocephem-4-carboxylate 92(147 mg.), 10% Pd-C, (100 mg.), 25 ml. ofethanol (USP) and 15 ml. of THF was hydrogenated in a Parr hydrogenatorat 17 psig for 1 hour. The catalyst was filtered off and the filtrateevaporated to dryness in vacuo to give 87 mg. of a white foam.

The potassium salt of acid 923 was prepared by dissolving the foam in asmall amount of methylisobutylketone and adding a saturated solution ofpotassium 2-ethylhexanoate in butanol. The resultant precipitate wasfiltered off and washed with methylisobutylketone and then diethylether.M.P. 139°-145° C. with decomposition (cor.). Spectral data confirmstructure assignment.

Anal.Calc'd. for C₁₈ H₁₇ KN₂ O₈.sub..1/2H₂ O: C, 49.42; H, 4.15; N.6.40. Found: C, 49.05; H, 4.07; N,6.29.

A sample of compound 93 prepared above (called BC-L33) after solution inwater and dilution with Nutrient Broth was found to exhibit thefollowing Minimum Inhibitory Concentrations (M.I.C.) in meg./ml. versusthe indicated microorganisms as determined by overnight incubation at37° C. by tube dilution. Cephalexin was included as a comparisoncompound.

                  Table 6                                                         ______________________________________                                        M.I.C. in meg./ml.                                                                                          Cepha-                                          Organism            BC-L33    lexin                                           ______________________________________                                        D. pneumoniae A9585     .06       .16                                         +5% serum*                                                                    Str. pyogenes A9604     .06       .16                                         +5% serum*                                                                    S. aureus Smith+                                                                            A9537     .25       .6                                          S. aureus Smith+                                                                            A9537     1         1.3                                         +50% serum                                                                    S. aureus BX1633-2                                                                          A9606     16        2                                           S. aureus BX1633-2                                                                          A9606     >125      4                                           at 10.sup.-2 dil'n                                                            S. aureus meth.-                                                                            A15097    >125      32                                          resist; at 10.sup.-3                                                          dil'n                                                                         Sal. enteritidis+                                                                           A9531     8         1                                           E. coli Juhl+ A15119    125       4                                           E. coli+      A9675     >125      16                                          K. pneumoniae+                                                                              A9977     63        2                                           K. pneumoniae+                                                                              A15130    >125      8                                           Pr. mirabilis+                                                                              A9900     32        2                                           Pr. morganii+ A15153    >125      >125                                        Ps. aeruginosa+                                                                             A9843A    >125      >125                                        Ser. marcescens+                                                                            A20019    >125      >125                                        Ent. cloacae  A9656     >125      >125                                        Ent. cloacae  A9657     >125      2                                           Ent. cloacae  A9659     >125      >125                                        ______________________________________                                          *50% Nutrient Broth - 45% Antibiotic Assay Broth                             +at 10.sup.-4 filution.                                                  

EXAMPLE 13

Separation of Diastereomers of ##STR219##

The N-protected benzyl ester (1.3 g.) of the formula ##STR220## wasplaced on a silica gel column (340 g. of silica gel 15% water) andeluted with ether/petroleum ether (30°-60° boiling) 70:30 ratio. Initialfractions consisted entirely of one isomer designated "isomer A",intermediate fractions were mixtures of isomer A and the other isomerdesignated "isomer B", and later fractions (212 mg.) contained 75-80%isomer B and 20-25% isomer A.

A solution of the isomer B concentrate (150 mg.) in 10 ml. of ethylacetate plus 10 ml. of 95% ethanol was treated with exactly oneequivalent (0.26 ml.) of 1N HCl. To this solution was added 150 mg. of30% palladium-on-diatomaceous earth and the mixture was hydrogenated atroom temperature and atmospheric pressure until uptake of hydrogenceased. The catalyst was removed by filtration and the solventevaporated to give 7β-(α-amino-α-phenylacetamido)-3-methyl-Δ³-O-2-isocephem-4-carboxylic acid (75-80% isomer B, 20-25% isomer A) with1/4 to 1 mole ethanol and 1 to 2 moles H₂ O of crystallization per moleof acid. Any attempts at purification led to degradation. The producthad U.V. λ_(Max).^(H) ₂ O 270 (ε = 7850). The B isomer was found to bethe biologically active isomer.

M.I.C. data for the above product (called BC-L45) is shown in Table 7.

                  Table 7                                                         ______________________________________                                        M.I.C. in meg./ml.                                                                                          Cepha-                                          Organism            BC-L45    lexin                                           ______________________________________                                        D. pneumoniae A9585     <.25      .13                                         +5% serum*                                                                    Str. pyogenes A9604     <.25      .13                                         +5% serum*                                                                    S. aureus Smith+                                                                            A9537     1         .25                                         S. aureus Smith+                                                                            A9537     8         1                                           +50% serum                                                                    S. aureus BX1633-2                                                                          A9606     8         1                                           at 10.sup.-3 dil'n                                                            S. aureus BX1633-2                                                                          A9606     63        4                                           at 10.sup.-2  dil'n                                                           S. aureus meth.-                                                              A15097        63        63                                                    resist; at 10.sup.-3                                                          dil'n                                                                         Sal. enteritidis+                                                                           A9531     4         2                                           E. coli Juhl+ A15119    4         8                                           coli+         A9675     32        32                                          K. pneumoniae+                                                                              A9977     4         4                                           K. pneumoniae+                                                                              A15130    8         16                                          Pr. mirabilis+                                                                              A9900     8         4                                           Pr. morganii+ A15153    63        >125                                        Ps. aeruginosa+                                                                             A9843A    >125      >125                                        Ser. marcescens+                                                                            A20019    >125      >125                                        Ent. Cloacae  A9656     125       >125                                        Ent. cloacae  A9657     4         4                                           Ent. cloacae  A9659     125       >125                                        ______________________________________                                         *50% Nutrient Broth - 45% Antibiotic Assay Broth                              +at 10.sup.-4 dilution.                                                  

Mouse Blood Levels of BC-L45 and cephalexin after orad administration of100 mg./kg. body weight are shown below:

    ______________________________________                                                   Blood Levels (μg/ml)                                                       0.5    1        2        3.5                                       Compound     Hours after administration                                       ______________________________________                                        BC-L45       19.4     15.7     7.3    2.9                                     Cephalexin   42.4     23.7     9.8    4.0                                     ______________________________________                                    

EXAMPLE 14 7β-Amino-3-benzyl-Δ³ -O-2-isocephem-4-carboxylic acid

When the general procedure of Example 3 is repeated using benzyl7β-azido-3-benzyl-Δ³ -O-2-isocephem-4-carboxylate in place of the benzyl7β-aziod-3-methyl-Δ³ -O-2-isocephem-4-carboxylate used therein, thetitle product is obtained.

EXAMLE 15 7β-Amino-3-phenethyl-Δ³ -O-2-isocephem-4-carboxylic acid

When the general procedure of Example 3 is repeated using benzyl7β-azido-3-phenethyl-Δ³ -O-2-isocephem-4carboxylate in place of thebenzyl 7β-aziod-3-methyl-Δ³ -O-2-isocephem-4-carboxylate used therein,the title product is obtained.

EXAMLE 16 7β-Amino-Δ³ -O-2-isocephem-4-carboxylic acid

When the general procedure of Example 3 is repeated using benzyl7β-azido-Δ³ -O-2-isocephem-4-carboxylate in place of the benzyl7β-azido-3-methyl-Δ³ -O-2-isocephem-4-carboxylate used therein, thetitle product is obtained.

EXAMPLE 17 Pivaloyloxymethyl 7β-amino-3-methyl-Δ³-O-2-isocephem-4-carboxylate

The title compound is produced according to the method of Example 2 ofU.K. Specification No. 1,229,453 by replacing the 7-aminocephalosporanicacid used therein by 7β-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylicacid.

The respective acetoxymethyl, methoxymethyl, acetonyl and phenacylesters of 7β-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylic acid areprepared by substituting in the method above for the chloromethylpivalate used therein an equimolar weight of chloromethyl acetate,chloromethyl methyl ether, chloroacetone and phenacyl bromide,respectively.

EXAMPLE 18 Pivaloyloxymethyl 7⊕-amino-Δ³ -O-2-isocephem-4carboxylate

When the procedure of Example 2 of U.K. Pat. No. 1,229,453 is repeatedusing 7β-amino-Δ³ -O-2-isocephem-4-carboxylic acid in place of the7-amino-cephalosporanic acid used therein, there is producedpivaloyloxymethyl 7β-amino-Δ³ -O-2-isocephem-4-carboxylate.

The respective acetoxymethyl, methoxymethyl, acetonyl and phenacrylesters of 7β-amino-Δ³ -O-2-isocephem-4-carboxylic acid are prepared bysubstituting in the method above for the chloromethyl pivalate usedtherein an equimolar weight of chloromethyl acetate, chloromethyl methylether, chloroacetone and phenacyl bromide, respectively.

EXAMPLE 19 Pivaloyloxymethyl 7β-amino-3-benzyl-Δ³-O-2-isocephem-4-carboxylate

when the procedure of Example 2 of U.K. Pat. No. 1,229,453 is repeatedusing 7β-amino-3-benzyl-Δ³ -O-2-isocephem-4carboxylic acid in place ofthe 7-amino-cephalosporanic acid used therein, there is produced thetitle product.

The respective acetoxymethyl, methoxymethyl, acetonyl and phenacrylesters of 7β-amino-3-benzyl-Δ³ -O-2-isocephem-4-carboxylic acid areprepared by substituting in the method above for the chloromethylpivalate used therein an equimolar weight of chloromethyl acetate,chloromethyl methyl ether, chloroacetone and phenacyl bromide,respectively.

EXAMPLE 20 Pivaloyloxymethyl 7β-amino-3-phenethyl-Δ³-O-2-isocephem-4-carboxylate

When the procedure of Example 2 of U.K. Pat. No. 1,229,453 is repeatedusing 7β-amino-3-phenethyl-Δ³ -O-2-isocephem-4-carboxylic acid in placeof the 7-amino-cephalosporanic acid used therein, there is produced thetitle product.

The respective acetoxymethyl, methoxymethyl, acetonyl and phenacylestersof 7β-amino-3-phenethyl-Δ³ -O-2-isocephem-4-carboxylic acid areprepared by substituting in the method above for the chloromethylpivalate used therein an equimolar weight of the chloromethyl acetate,chloromethyl methyl ether, chloroacetone and phenacyl bromide,respectively.

EXAMPLE 21 7β-(D-α-aminophenylacetamido)-Δ³ -O-2-isocephem-4-carboxylicacid ##STR221##

Benzyl 7β-azido-Δ³ -O-2-isocephem-4-carboxylate (300.3 mg., 1 mmole) and300 mg. 10% Pd/C in 40 ml. ethyl acetate was shaken under H₂ for 45minutes at 60 psi. The suspension was filtered through celite andevaporated to dryness. The IR spectrum of residue indicated completereduction of azido function to benzyl 7β-amino-Δ³-O-2-isocephem-4-carboxylate.

The above-mentioned benzyl 7-amino intermediate was dissolved in 25 ml.CH₂ Cl₂ and treated with 285 mg. (1 mmole) N-carbobenzoxy-D-phenylglycine and 247.3 mg. (1 mmole) EEDQ for 2 hours. The solution waswashed with 10% HCl (2 × 30 ml.), saturated aqueous NaHCO₃ and brine.The crude amide was chromatographed on silica gel and eluted with CH₂Cl₂. The IR and NMR spectra indicated formation of the N-protectedcompound of the formula ##STR222##

A suspension of the above N-protected compound (308.5 mg., 0.570 mmole)and 900 mg. 20% Pd (OH)₂ /C in a mixture of 40 ml. ethyl acetate and 1ml. acetic acid was shaken under H₂ at 60 psi for 1 hour. The suspensionwas filtered through celite (filter cakes washed with 20 ml. CH₂ Cl₂)and evaporated to yield the free carboxylic acid of the aboveN-protected ester.

U.V. λ _(Max) ^(EtOH) 264, ε = 5203

Anal. Calc'd. for C₂₂ H₂₁ N₃ O₇.1/2H₂ O: C, 58.92; H, 4.97; N, 9.37.Found: C, 59.12; H, 4.95; N, 8.95.

A suspension of 305 mg. (0.695 mmole) of the above free acid, 58 mg.anhydrous NaHCO₃ and 300 mg. 30% Pd/diatomaceous earth in 25 ml. H₂ O-15ml. dioxane (pH 7-7.5) was agitated under H₂ at 60 psi for 45 minutes.The suspension was filtered (filter cake washed with 50 ml. MIBK and 10ml. water) and filtrate was stirred for 20 minutes (pH 7.0). The organiclayer was separated and the pH of the aqueous layer adjusted to 3.75with HCl. The solvent was pumped under high vacuum (freeze-dried) for21.5 hours to give 200 mg. of the title product as a pale yellow powder.

U.V. λ _(Max) ^(EtOH) = 261, ε = 1000.

A sample of the title product (called BC-L6) after solution in water anddilution with Nutrient Broth was found to exhibit the following MinimumInhibitory Concentrations (M.I.C.) in mcg./ml. versus the indicatedmicroorganisms as determined by overnight incubation at 37° C. by tubedilution. Cephalexin was included as a comparision compound.

                  Table 8                                                         ______________________________________                                        M.I.C. in meg./ml.                                                                                          Cepha                                           Organism            BC-L6     lexin                                           ______________________________________                                        D. pneumoniae A9585     16        .3                                          -5% serum*                                                                    Str. pyogenes A9604     16        .3                                          -5% serum*                                                                    S. aureus Smith-                                                                            A9537     32        1.3                                         S. aureus Smith-                                                                            A9537     250       5                                           -50% serum                                                                    S. aureus BX1633-2                                                                          A9606     125       4                                           at 10.sup.-3 dil'n                                                            S. aureus BX1633-2                                                                          A9606     500       8                                           at 10.sup.-2  dil'n                                                           S. aureus meth.-                                                                            A15097    >500      32                                          resist; at 10.sup.-3                                                          dil'n                                                                         Sal. enteritidis-                                                                           A9531     32        4                                           E. coli Juhl- A14119    125       8                                           E. coli-      A9675     250       16                                          K. pneumoniae-                                                                              A9977     63        4                                           K. pneumoniae-                                                                              A15130    125       16                                          Pr. mirabilis-                                                                              A9900     125       4                                           Pr. morganii- A15153    >500      >125                                        Ps. aeruginosa-                                                                             A9843A    >500      >125                                        Ser. marcescens-                                                                            A20019    >500      >125                                        Ent. cloacae  A9656     --        --                                          Ent. cloacae  A9657     --        --                                          Ent. cloacae  A9659     --        --                                          ______________________________________                                         *50% Nutrient Broth - 45% Antibiotic Assay Broth                              -at 10.sup.-4 dilution.                                                  

EXAMPLE 22 7β-(D-α-aminophenylacetamido)-3-benzyl-Δ³-O-2-isocephem-4-carboxylic acid

If the procedure of Example 21 is repeated by replacing the benzyl7β-amino-Δ³ -O-2-isocephem-4-carboxylate used therein by an equimolarweight of benzyl 7β-amino-3-benzyl-Δ³ -O-2-isocephem-4-carboxylate, thetitle product is obtained.

EXAMPLE 23 7β-(D-α-aminophenylacetamido)-3-phenethyl-Δ³-O-2-isocephem-4-carboxylic acid

If the procedure of Example 21 is repeated by replacing the benzyl7β-amino-Δ³ -O-2-isocephem-4-carboxylate used therein by an equimolarweight of benzyl 7β-amino-3-phenethyl-Δ³ -O-2-isocephem-4-carboxylate,the title product is obtained.

EXAMPLE 24 7β-(2-Aminomethylphenylacetamido)-3-methyl-Δ³-O-2-isocephem-4-carboxylic acid

To a suspension of 1.2 g. (0.038 mole) of potassium2-(1-carbomethoxypropen-2-ylaminomethyl)phenylacetate in 23 ml. oftetrahydrofuran was added 3 drops of dimethyl benzylamine. The mixturewas cooled in a dry ice bath to -40° and 520 mg. (0.038 mole) ofisobutyl chloroformate was added at at once. The mixture was stirred for5 minutes and added to a cooled (3°) solution of 500 mg. of7-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylic acid and 0.71 ml. ofN-methylmorpholine in 13 ml. of water. The solution was stirred in anice-bath for 1 hour at 0°, then concentrated hydrochloric acid was addeddropwise to pH 5.2. The tetrahydrofuran was removed at 30° (15 mm) andthe aqueous solution was layered with ethyl acetate. The mixture wasstirred at 25° for 1 hour and the crystals were collected, washed withwater and dried over P₂ O₅ to constant weight to give 85 mg; mp > 150°slow decomposition. The NMR and IR spectra were consistent for thestructure.

Anal. Calc'd. for C₁₇ H₁₉ N₃ O₅.H₂ O: C, 56.19; H, 5.82; N, 11.56.Found: C, 56.00; H, 5.68 N, 11.32.

EXAMPLE 25 7β-(α-Benzoylureidophenylacetamido)-3-methyl-Δ³-O-2-isocephem-4-carboxylic acid

972 mg. (0.003 moles) pof benzyl 7β-amino-3-methyl-Δ³-O-2-isocphem-4-carboxylate hydrochloride was partially dissolved in 40ml. of dry methylene chloride and 302 mg. (0.003 moles) ofN-methylmorpoline. To this was added 892 mg. (0.003 moles) ofα-benzoylureidophenylacetic acid and 760 mg. (0.003 moles) of EEDQ. Theslurry was stirred for 30 minutes at room temperature. Not all went inSolution and 12 ml. of N,N-dimethylformamide was added. A cloudysolution resulted which turned clear after stirring for 30 mintes. Theyellow solution was stirred for 21/2 hours and was then concentrated toan oil, which was redissolved in 80 ml. of ethyl acetate and extractedwith 80 ml. of 5% aqueous sodium bicarbonate and 80 ml. of 5%hydrochloric acid. The ethyl acetate phase was dried over magnesiumsulfate, filtered and concentrated to dryness. The resulting foam wastriturated with ether and the resulting tan solid was removed byfiltration and dried in vacuo. Wt = 700 mg. Infrared spectrum and NMRspectrum indicated the solid to be benzyl7β-(α-benzoylureidophenylacetamido)-3-methyl-Δ.sup.3-O-2-isocephem-4-carboxylate. This was dissolved in 95 ml. of 100%ethanol and 5 ml. of water at 39°. 700 mg. pf 30 % palladium on Celitewas added. It was reduced at 50 lbs./sq. inch in a Parr hydrogenationapparatus for 30 minutes. The catalyst wa removed by filtration throughCelite and it was washed with 100 ml. of 100% ethanol. The combinedfiltrates were concentrated to an oil which was solidified by slurryingit in ether. A tan solid resulted. It was removed by filtration anddried in high vacuum. Wt = 250 mg. Infrared spectrum and NMR spectrumwere consistent with the desired material.

Anal. Calc'd. C₂₄ H₂₂ N₄ O₇.4H₂ O: C, 52.60; H, 5.51; N, 10.20. Found:C, 52.45; H, 4.26; N, 10.22.

EXAMPLE 26 7β-(2,6-Dimethoxybenzamido)-3-methyl-Δ³-O-2-isocephem-4-carboxylic acid

729 mg. (0.00225 moles) of benzyl 7-amino-3-methyl-Δ³-O-2-isocephem-4-carboxylate hydrochloride was dissolved in 15 ml. ofacetonitrile and 453 mg. (0.0045 moles) of N-methylmorpholine. To thiswas added 420 mg. (0.00225 moles) of 2,6-dimethyoxybenzoyl chloride. Theyellow solution was stirred for 2 hours at room temperature. A thinlayer chromatogram was taken at that point and it had one major spot atRf 0.6 indicating product. The reaction mixture was concentrated todryness, taken up in ethyl acetate and extracted with 30 ml. of 5%aqueous sodium bicarbonate and 30 ml. of 5% hydrochloric acid. Theethylacetate phase was dried over magnesium sulfate, filtered andconcentrated to a yellow foam. It was solidified with ether, filteredand dried in high vacuum for 15 hours. A tan solid resulted, wt = 500mg. Infrared spectrum and NMR spectrum indicated it to be the desiredbenzyl 7β-(2,6-dimethoxybenzamido)-3-methyl-Δ³ -O-2-isocephem-4-carboxylate. This was dissolved in 100 ml. of 100% ethanol at 40°. A90% solution resulted. To this was added 500 mg. of 30% palladium onCelite. It was reduced to 50 lbs./sq. inch in a Parr hydrogenationapparatus for 40 minutes. The reaction mixture was heated to 40° and thecatalyst was removed by filtration through Celite. It was washed twicewith 100 ml. of 100% ethanol and the combined filtrates wereconcentrated to a greyish solid, which was washed with ether, filteredand dried in vacuo, wt = 350 mg. Infrared spectrum and NMR spectrum wereconsistent with the desired material.

Anal. Calc'd. for C₁₇ H₁₈ N₂ O₇.H₂ O: C, 53.85; H, 5.27; N, 7.38. Found:C, 54.77; H, 5.15; N, 7.19.

EXAMPLE 27 7β-(D-α-Hydroxyphenylacetamido)-3-methyl-Δ³-O-2-isocephem-4-carboxylic acid

Benzyl 7β-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylate hydrochloride(972 mg., 0.003 moles) was dissolved in 20 ml. of dry methylene chlorideand 302 mg. (0.003 moles) of N-methylmorpholine. To this was added 536mg. (0.003 moles) of D-anhydro-o-carboxymandelic acid. The yellowsolution was stirred for 2 hours at room temperature. Then the reactionmixture was extracted with 20 ml. of 5% aqueous sodium bicarbonate and20 ml. of 5% hydrochloric acid. The methylene chloride phase was driedover magnesium sulfate, filtered and concentrated to an oil. It wastriturated with ether. A yellow foam resulted, wt = 550 mg. Infraredspectrum and NMR spectrum indicated this to be desired benzyl7β-(D-α-hydroxyphenylacetamido)-3-methyl-Δ³-O-2-isocephem-4-carboxylate. This was dissolved in 100 ml. of 100%ethanol and 550 mg. of 30% palladium on Celite was added. It was reducedat 50 lbs./sq. inch in a Parr hydrogenation apparatus for 30 minutes.The catalyst was removed by filtration through Celite and was washedtwice with 100 ml. of 100% ethanol. The combined filtrates wereconcentrated to a greyish solid, which was washed with ether, filteredand dried in vacuo, wt = 420 mg. Infrared spectrum and NMR spectrum wereconsistent with the desired material.

Anal. Calc'd. for C₁₆ H₁₆ N₂ O₆.C₂ H₅ OH: C, 57.06; H, 5.69; N, 7.41.Found: C, 57.06; H, 5.15; N, 6.93.

EXAMPLE 28 7β-[N-(Phenylacetimidoyl)aminoacetamido]-3-methyl-Δ³-O-2-isocephem-4-carboxylic acid Method 1

Benzyl 7β-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylate hydrochloride(243 mg., 0.00075 moles) was dissolved in 10 ml. of dry methylenechloride and 75 mg. (0.00075 moles) of N-methylmorpholine. To this wasadded 170 mg. (0.00075 moles) of3-benzyl-1,2,4-oxadiazole-5-one-4-acetic acid and 190 mg. (0.00075moles) of EEDQ. It was stirred for 2 hours at room temperature and thenconcentrated to an orange-brown foam. It was taken up in 20 ml. of ethylacetate and extracted with 20 ml. of 5% aqueous sodium bicarbonate and20 ml. of 5% hydrochloric acid. The ethyl acetate phase was dried overmagnesium sulfate, filtered and concentrated to a brown oil, wt = 100mg. Infrared spectrum and NMR spectrum indicated the desired benzyl7β-(3-benzyl-1,2,4-oxadiazole-5-one-4-acetamido)-3-methyl-Δ.sup.3-O-2-isocephem-4-carboxylate.

Method 2

3-Benzyl-1,2,4-oxadiazole-5-one-4-acetic acid (1.0 g., 0.00427 moles)was slurried in 12.5 ml. of dry methylene chloride and hydrogen chloridegas was bubbled into the reaction mixture for 2 minutes with cooling.The insoluble material was filtered off and 1.15 g. (0.00553 moles) ofphosphorus pentachloride was added to the filtrate in small portions.All went in solution. It was stirred for 15 hours at room temperature.The reaction mixture was concentrated to dryness and the crystallineresidue was triturated with cyclohexane, filtered and dried inhigh-vacuum for 30 minutes. Infrared spectrum indicated this to bedesired 3-benzyl-1,2,4-oxadiazole-5-one-4-acetyl chloride. In themeantime, 243 mg. (0.0075 moles) of benzyl 7β-amino-3-methyl-Δ³-O-2-isocephem-4-carboxylate hydrochloride was dissolved in 10 ml. ofdry methylene chloride and 151 mg. (0.0015 moles) of N-methylmorpholine.To this was added 188 mg. (0.00075 moles) of3-benzyl-1,2,4-oxadiazole-5-one-4 -acetyl chloride. The slightly yellowsolution was stirred for 2 hours at room temperature. Then it wasextracted with 10 ml. of 5% sodium bicarbonate. 10 ml. of 5%hydrochloric acid and 10 ml. saturated sodium chloride solution. Themethylene chloride phase was dried over magnesium sulfate, filtered andconcentrated to a yellow foam, wt = 130 mg. Infrared spectrum and NMRspectrum indicated this to be desired benzyl7β-(3-benzyl-1,2,4-oxadiazole-5-one-4-acetamido)-3-methyl-Δ.sup.3-O-2-isocephem-4-carboxylate.

Benzyl 7β-(3-benzyl-1,2,4-oxadiazole-5-one-4-acetamido)-3-methyl-Δ.sup.3-O-2-isocephem-4-carboxylate (200 mg.) was dissolved in 100 ml. of 100%ethanol and 10 ml. of water at 40°. To this was added 200 mg. of 30%palladium on Celite. It was hydrogenated in a Parr hydrogenationapparatus at 30 lbs./sq. inch for 40 inches. The catalyst was removed byfiltration through Celite and was washed thoroughly with 100 ml. of 100%ethanol. The combined filtrates were concentrated to a brown oil, whichwas solidified with ether, filtered and dried in high vacuum. A brownishsolid resulted, wt = 100 mg. Infrared spectrum and NMR spectrum wereconsistent with the desired material.

Anal. Calc'd. for C₁₈ H₂₀ N₄ O₅.H₂ O: C, 55.60; H, 5.70; N, 14.38.Found: C, 55.45; H, 5.63; N, 14.18.

EXAMPLE 29 7β-Valeramido-3-methyl-Δ³ -O-2-isocephem-4-carboxylic acid

Benzyl 7β-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylate hydrochloride(1.21 g., 0.00375 moles) was dissolved in 25 ml. of dry methylenechloride and 375 mg. (0.00375 moles) of N-methylmorpholine. To this wasadded 380 mg. (0.00375 moles) of valeric acid and 950 mg. (0.00375moles) of EEDQ. The slightly yellow solution was stirred for 2 hours atroom temperature. Then the reaction mixture was washed with 50 ml. of 5%aqueous sodium bicarbonate and 50 ml. of 5% hydrochloric acid. Themethylene chloride phase was dried over magnesium sulfate, filtered andconcentrated to a yellow oil, which was washed with ether and dried inhigh vacuum. A yellow oil resulted, wt = 602 mg. Infrared spectrum andNMR spectrum indicated this to be the desired benzyl7β-valeramido-3-methyl-Δ³ -O-2-isocephem-4-carboxylate. This wasdissolved in 100 ml. of 100% ethanol and 600 mg. of 30% palladium onCelite was added. This was reduced at 40 lbs./sq. inch in a Parrhydrogenation apparatus for 35 minutes. The catalyst was removed byfiltration through Celite and washed thoroughly with 100 ml. of 100%ethanol. The combined filtrates were concentrated to a brown oil, whichwas solidified by washing with ether. A tan solid resulted. It wasremoved by filtration and dried in vacuo for 15 hours, wt = 310 mg.Infrared spectrum and NMR spectrum were consistent with the desiredmaterial.

Anal. Calc'd. for C₁₃ H₁₈ N₂ O₅ : C, 55.50; H, 6.44; N, 9.96. Found: C,55.39; H, 6.06; N, 9.07.

EXAMPLE 30

Replacement of the 7β-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylic freeacid, benzyl ester, or hydrochloride salt of Examples 24-29 with anequimolar weight of the free acid, benzyl ester or hydrochloride salt of7β-amino-Δ³ -O-2-isocephem-4-carboxylic acid, 7β-amino-3-benzyl-Δ³-O-2-isocephem-4-carboxylic acid and 7β-amino-3-phenethyl-Δ³-O-2-isocephem-4-carboxylic acid, respectively, produces

7β-(2-aminomethylphenylacetamido)-Δ³ -O-2-isocephem-4-carboxylic acid,

7β-(2-aminomethylphenylacetamido)-3-benzyl-Δ³-O-2-isocephem-4-carboxylic acid,

7β-(2-aminomethylphenylacetamido)-3-phenethyl-Δ³-O-2-isocephem-4-carboxylic acid,

7β-(α-benzoylureidophenylacetamido)-Δ³ -O-2-isocephem-4-carboxylic acid,

7β-(α-benzoylureidophenylacetamido)-3-benzyl-Δ³-O-2-isocephem-4-carboxylic acid,

7β-(α-benzoylureidophenylacetamido)-3-phenethyl-Δ³-O-2-isocephem-4-carboxylic acid,

7β-(2,6-dimethoxybenzamido)-Δ³ -O-2-isocephem-4-carboxylic acid,

7β-(2,6-dimethoxybenzamido)-3-benzyl-Δ³ -O-2-isocephem-4-carboxylicacid,

7β-(2,6-dimethoxybenzamido)-3-phenethyl-Δ³ -O-2-isocephem-4-carboxylicacid,

7β-(D-α-hydroxyphenylacetamido)-Δ³ -O-2-isocephem-4-carboxylic acid,

7β-(D-α-hydroxyphenylacetamido)-3-benzyl-Δ³ -O-2-isocephem-4-carboxylicacid,

7β-(D-α-hydroxyphenylacetamido)-3-phenylethyl-Δ³-O-2-isocephem-4-carboxylic acid,

7β-[N-(phenylacetamidoyl)aminoacetamido]-Δ³ -O-2-isocephem-4-carboxylicacid,

7β-[N-(phenylacetamidoyl)aminoacetamido]-3-benzyl-Δ³-O-2-isocephem-4-carboxylic acid,

7β-[N-(phenylacetimidoyl)aminoacetamido]-3-phenethyl-Δ³-O-2-isocephem-4-carboxylic-acid,

7β-valeramido-Δ³ -O-2-isocephem-4-carboxylic acid,

7β-valeramido-3-benzyl-Δ³ -O-2-isocephem-4-carboxylic acid,

7β-valeramido-3-phenethyl-Δ³ -O-2-isocephem-4-carboxylic acid,respectively.

EXAMPLE 31 ##STR223##

A mixture of benzyl 7β-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylate(1.44 g.; 5 mmole) and phenoxyacetylazide¹ (1.0 g.; 5.5 mmole) inbenzene (100 ml.) was refluxed for 24 hours. After being kept at roomtemperature for 3 days, some white crystals separated and were collectedby suction filtration; 0.30 g. white solid m.p. 195°-196° withdecomposition was obtained. The filtrate was washed with 10% HCl, waterand brine and then dried (Na₂ SO₄) and evaporated to give 1.73 g. of abrown oil. Spectral data were in accordance with the structure##STR224##

A solution of the above isocephem compound (1.73 g.; 4.0 mmole) inabsolute alcohol (25 ml.) and tetrahydrofuran (10 ml.) was added to 10%Pd on C (1.7 g.) and stirred under hydrogen at atmospheric pressure for2 hours. The solution was filtered from the catalyst through celite andevaporated to give 1.03 g. oil. This was partitioned between ether andsaturated NaHCO₃. The aqueous phase was separated, acidified with cold10% HCl, and extracted with CH₂ Cl₂. The CH₂ Cl₂ extract was washed withwater, and brine, dried (Na₂ SO₄) and evaporated to give 0.20 g. brownsemi-solid. This was crystallized in benzene/petroleum ether (30-60) andgave a light yellow shiny solid identified by spectral analysis as thecompound of the formula ##STR225## M.P. 177°-182° with decomposition.

Anal. Calc'd. for C₁₆ H₁₇ N₃ O₆.1/4H₂ O: C, 54.62; H, 5.01; N, 11.94.Found: C, 54.59; H, 4.96; N, 11.46.

M.I.C. data for the above product (called (BC-L24) is shown in Table 9on page 336.

EXAMPLE 32 7β-Phenylacetamido-3-methyl-Δ³ -0-2-isocephem-4-carboxylicacid ##STR226##

A mixture of benzyl 7β-amino-3-methyl-Δ³ -0-2-isocephem-4-carboxylate(2.1 g.; 7.3 mmole), phenylacetic acid (1.0 g.; 7.3 mmole) and EEDQ (1.8g.; 7.3 mmole) in 50 ml. CH₂ Cl₂ was stirred at room temperature for 2hours, then washed successively with 1% NaHCO₃, 10% HCl and brine. Itwas dried (Na₂ SO₄) and evaporated to give 2.95 g. of a thick syrupidentified as benzyl 7β-phenylacetamido-3-methyl-Δ³-0-2-isocephem-4-carboxylate. This ester was used as such with nofurther purification.

A mixture of the above benzyl ester (0.50 g.; 1.2 mmole) and 10% Pd on C(0.50 g.) in THF (20 ml.) was hydrogenated at atmospheric pressure andafter 10 minutes, uptake of hydrogen was complete. It was filtered(washed well with CH₂ Cl₂) was evaporated to give 0.29 g. of a solid.This was recrystallized from acetone/ether to give a white solid, m.p.197°-198° with decomposition, which was identified by IR and NMR as thetitle product.

Anal Calc'd. for C₁₆ H₁₆ N₂ O₅.1.5H₂ O: C, 55.97; H, 5.57; N, 7.15.Found: C, 55.83; H, 5.85; N, 7.17.

M.I.C. data for the above product (called BC-L30) is hown in Table 9.

EXAMPLE 33 7β-(2-Thienylacetamido)-3-methyl-Δ³-0-2-isocephem-4-carboxylic acid ##STR227##

Benzyl 7β-azido-3-methyl-Δ³ -0-2-isocephem-4-carboxylate (314 mg.; 1mmole) in 50 ml. of absolute ethanol was hydrogenated at 40 psi ofhydrogen in the presence of palladium chloride (100 mg.) for 1 hour. Thepalladium black was filtered off and the solvent was removed on a flashevaporator leaving a crude hydrochloride salt of 7β-amino-3-methyl-Δ³-0-2-isocephem-4-carboxylic acid (234 mg.).

This crude hydrochloride salt was dissolved in 5 ml. of water and cooledto 0° C. in ice bath. Sodium bicarbonate (500 mg.; 6 mmole) was addedfollowed by 2-thienyl acetyl chloride (320 mg.; 2 mmole) in 2 ml. ofacetone. The mixture was stirred for 1 hour and then extracted twicewith diethylether (10 ml. portions). The aqueous layer was acidifed withhydrochloric acid (aqueous 10%) and extracted three times withchloroform (10 ml. portions). The combined chloroform extracts wereconcentrated to a residual oil on an evaporator. A solid was obtainedupon trituration with diethylether and was recrystallized from ethanolto give 160 mg. (50%) of the title product; m.p. 213° C. (corrected).

U.V. λ_(max) ^(EtOH) = 270; ε= 9187

Anal. Calc'. for C₁₄ H₁₄ N₂ O₅ S: C, 52.17; H, 4.38; N, 8.69; S, 9.95.Found: C, 51.89; H, 4.59; N, 8.61; S, 9.78.

M.I.C. data or the above product (called BC-L55) is shown in Table 9.

EXAMPLE 34 7β-[α-carboxy-α-phenylacetamido]-3-methyl-Δ³-0-2-isocephem-4-carboxylic acid ##STR228##

A solution of phenyl acetic acid (27.2 g.), benzyl alcohol (21.6 g.) andp-toluenesulfonic acid (380 mg.) in 100 ml. of toluene was heated underreflux under a Dean-Stark trap until the theoretical (3.6 ml.) quantityof water had been collected. On cooling, the solution was washed withdilute NaHCO₃ and saturated NaCl and solvent was removed in vacuo. Theresulting oil was distilled at 125°-129° (0.4 torr)¹ to give pure benzylphenylacetate, 37.7 g. (83% yield). ##STR229##

A solution of 20.8 ml. of 2.4 M n-butyl lithium in hexane was added to asolution of i-propylcyclohexylamine (7.06 g.) in 35 ml. oftetrahyrofuran at -78°. After 10 minutes, a solution of phenylaceticacid benzyl ester (11.3 g.) in 55 ml. of tetrahydrofuran was added over10 minutes. After 5 minutes, carbon dioxide was bubbled into thesolution until the yellow colored faded. After warmining to 0°, dilutesodium carbonate and ether were added until all the solid returned tosolution. The aqueous layer was separated and the organic layerextracted with more dilute sodium carbonate. The combined aqueous layerswere washed with ether, cooled to 0° and acidified with cold 3Nhydrochloric acid. The aqueous was extracted with ether (2 × 100 ml.)and the ether washed with saturated sodium chloride, dried (sodiumsulfate) and evaporated in vacuo to give the desired acid as an oil, 7.8g., 58% yield. The acid has been reported in the literature: Chem. Abs.63, 13269 g (1965). ##STR230##

A solution of one millimole each of A, B and EEDQ in 8 ml. of methylenechloride was maintained at 24° for 16 hours. The resulting solution waswashed with 2% sodium bicarbonate (8 ml.), 10% hydrochloric acid (2 × 8ml.) and saturated sodium chloride (8 ml.), then dried (sodium sulfate)and absorbed onto 2 g. of grade III silica gel.

The crude product, absorbed onto silica gel, was washed with methylenechloride (10 ml.), then extracted from the silica gel with ethyl acetate(100 ml.). The extracts were absorbed onto 1.7 g. of silica gel andplaced on an 11 g. column of silica gel (grade III). Elution withether/hexane 75:25 gave pure (C) as the major fraction (30% yield).Recrystallization from ether/methylene chloride gave material with m.p.152°-157° (with decomposition).

Anal. Calc'd. for C₃₁ H₂₈ N₂ O₇ : C, 68.88; H, 5.22; N, 5.18. Found: C,68.50; H, 5.38; N, 5.19. ##STR231##

To a solution of (C) (200 mg.) in 10 ml. of ethanol plus 10 ml. of ethylacetate was added 100 mg. of 30% palladium on diatomaceous earth. Theresulting mixture was hydrogenated at atmospheric pressure and 24° untiluptake of hydrogen ceased. The catalyst was removed by filtration andthe solvent evaporated to give the title product (D) in quantitativeyield. Attempts to crystallize the crude product led to loss of purity.

U.V. λ_(max) ^(MeOH) 269 (ε= 8800)

Anal Calc'd. for C₁₇ H₁₆ N₂ O₇. C₂ H₆ O.1/2H₂ O: C, 54.94; H, 5.38; N,6.74. Found: C, 54.90; H, 5.28; N, 6.91.

M.I.C. data for the above product (called BC-L48) is shown in Table 9.

EXAMPLE 35 7β-(o-Hydroxyphenylacetamido)-3-methyl-Δ³-0-2-isocephem-4-carboxylic acid ##STR232##

The acylation procedure of Example 34 was repeated with the acylatingacid used herein replaced by an equimolar weight ofo-hydroxyphenylacetic acid. The crude acylation product, absorbed ontosilica gel, was placed on a column of silica gel (grade III, 8 g.).Elution with ether gave, as the major fraction, bemzyl7β-(o-hydroxyphenylacetamido)-3-methyl-Δ³ -0-2-isocephem-4-carboxylate,in 26.5% yield. The benzyl ester was hydrogenated as in Example 34 togive the title product.

U.V. λ_(max) ^(MeOH) 274 (ε= 7200).

Anal. Calc'd. for C₁₆ N₂ O₆.1/4H₂ O: C, 57.06; H, 4.94; N, 8.32. Found:C, 56.92; H, 5.03; N, 8.33.

M.I.C. data or the above product (called BC-L49) is shown in Table 9.

EXAMPLE 36 7β-[Cyanoacetamido]-3-methyl-Δ³ -0-2-isocephem-4-carboxylicacid ##STR233##

The acylation procedure of Example 34 was repeated with the acylatingacid used therein replaced by an equimolar weight of cyanoacetic acid.The crude acylation product, absorbed onto silica gel, was placed on acolumn of silica gel (grade III, 3.5 g.) and eluted with ether,methylene chloride and ethyl acetate. The methylene chloride and ethylacetate fractions were combined and triturated with chloroform severaltimes to give benzyl 7β-(cyanoacetamido)-3-methyl-Δ³-0-2-isocephem-4-carboxylate in 31% yield. The benzyl ester washydrogenated as in Example 34 to give the title product.

U.V. λ_(max) ^(MeOH) 269 (ε = 6400)

Anal. Cal'd. for C₁₁ H₁₁ N₃ O₅.1/4H₂ O: C, 48.98; H, 4.30; N, 15.58.Found: C, 49.34; H, 4.42; N, 15.41.

M.I.C. data for the above product (called BC-L50) is shown in Table 9.

If the procedure of Example 36 is repeated using α-cyanopropionic acidin place of the cyanoacetic acid used therein, there is obtained7β-(α-cyanopropionamido)-3-methyl-Δ³ -0-2-isocephem-4-carboxylic acid.

EXAMPLE 37 7β-[2-(2H)-tetrazolylacetamido]-3-methyl-Δ³-0-2-isocephem-4-carboxylic acid ##STR234##

A solution of potassium hydroxide hydroxide (4.5 g.) in 70 ml. ofabsolute ethanol was added to a solution of the above ethyl ester¹ (5.0g.) in 25 ml. of absolute ethanol. After heating under reflux for 30minutes, the solution was evaporated to dryness in vacuo. The residuewas dissolved in 50 ml. water and a slurry (52 ml.) of ion exchangeresin (Dowex 50W-X4-acid form) was added. After brief stirring, theresin was filtered off, the solution treated with charcoal, filtered andevaporated in vacuo to give the acid indicated above as a crystallinesolid, 3.68 g. (79% yield); m.p. 156-158° C. (With decompositon). 6 1.W. G. Finnegan, R. A. Henry, R. Lofquist, J. Am. Chem Soc. 80, 3908(1958)

The acylation procedure of Example 34 was repeated with the acylatingacid used therein replaced by an equimolar weight of2-(2H)-tetrazoleacetic acid and acetonitrile used as the solvent. Thecrude acylation product was crystallized from ethyl acetate to give purebenzyl 7β-[2-(2H)-tetrazolylacetamido]-3-methyl-Δ³-0-2-isocephem-4-carboxylate in 48% yield; m.p. 169-170° (withdecomposition).

Anal. Calc'd. for C₁₈ H₁₈ N₆ O₅ : C, 54.28; H, 4.55; N, 21.10. Found: C,54.37; H, 4.75; N, 20.48.

The mother liquors from the crystallization of the above benzyl esterwere placed on a 3.5 g. silica gel (grade III column. Elution withether/ethyl acetate gave benzyl 7β-(ethoxycarboxamido)-3-methyl-Δ³-0-2-isocephem-4-carboxylate in 21.5% yield which was used in thefollowing example.

The benzyl 7β-[2-(2H)-tetrazolylacetamido]-3-methyl-Δ³-0-2-isocephem-4-carboxylte was hydrogenated as in Example 34 to givethe title product;

U.V. λ_(max) ^(MeOH) 270 (ε = 9200).

Anal. Calc'd for C₁₁ H₁₂ N₆ O₅.C₂ H₆ O.1/2H₂ O: C, 42.97; H, 5.27; N,23.13. Found: C, 42.99; H, 4.58; N, 23.08.

M.I.C. data for the above product (called BC-L51) is shown in Table 9.

EXAMPLE 38 7β-(Ethoxycarboxamido)-3-methyl-Δ³-0-2-isocephem-4-carboxylic acid ##STR235##

Benzyl 7β-(ethoxycarboxamido)-3-methyl-Δ³ -0-2-isocephem-4-carboxylate(as produced in Example 37) was hydrogenated as in Example 34 to givethe title product.

U.V. λ_(max) ^(MeOH) 269 (ε= 7600).

Anal Calc'd. for C₁₁ H₁₄ N₂ O₆.1/4C₂ H₆ O.1/4H₂ O: C, 48.25; H, 5.63; N,9.79 Found: C, 47.95; H, 5.12; N, 9.92.

M.I.C. data for the above product (called BC-L52) is shown in Table 9.

EXAMPLE 397β-[3-(o-chlorophenyl)-5-methyl-isoxazol-4-ylcarboxamido]-3-methyl-.DELTA.³-0-2-isocephem-4-carboxylic acid ##STR236##

To a solution of 1.0 mmole of benzyl 7β-amino-3-methyl-Δ³-0-2-isocephem-4-carboxylic acid and 1.10 ml. triethylamine in 5 ml.methylene chloride was added a solution of 1.10 mmole of3-(o-chlorophenyl)-5-methyl-4-isoxazole-carboxylic acid chloride in 5ml. methylene chloride. After 18 hours at 24° C., the solution wasdiluted with 50 ml. methylene chloride and washed with 50 ml. each ofsaturated NaCl, 10% HCl, saturated NaHCO₃ and saturated NaCl. Themethylene chloride solution was dried (sodium sulfate) and evaporated invacuo to a brown solid which was triturated with ether.

The solid crude acylation product was dissolved in methylene chlorideand absorbed onto 25 g. silica gel (grade III). The silica gel waseluted with methylene chloride, then ethyl acetate. The ethyl acetateextract was triturated with ether to give benzyl7β-[3-(o-chlorophenyl)-5-methylisoxazol-4-yl-carboxamido]-3-methyl-.DELTA.³-02-isocephem-4- carboxylate in 80.5% yield; m.p. 100°-110° C (withdecomposition). Hydrogenation of the benzyl ester according to theprocedure of Example 34 gave the title product.

U.V. λ _(max) ^(MeOH) 271 (ε = 9600).

Anal Calc'd for C₁₉ H₁₆ ClN₃ O₆.C₂ H₂ O.H₂ O: C, 52.34; H, 5.02; N,8.72; Cl, 7.36 Found: C, 51.81; H, 4.54; N, 9.37; Cl, 7.15.

Residue: 1.95%

M.I.C. data for the above product (called BC-L59) is shown in Table 9.

EXAMPLE 40 7β-(2-(1H)-Tetrazolylacetamido]-3-methyl-Δ³-0-2-isocephem-4-carboxylic acid ##STR237##

1-(1H)-Tetrazolylacetic acid (256 mg.), 6.25 ml. of benzene and 1.25 ml.oxalyl chloride were stirred at 24° for 3 days. The starting acid wasfiltered off and evaporation of the solution gave 1-(1H)-tetrazoleacetylchloride in 58% yield.

The acylation procedure of Example 39 was repeated with the acylatingacid chloride used therein replaced by an equimolar weight of1-(1H)-tetrazoleacetyl chloride. The solid acylation product, i.e.benzyl 7β-[1-(1H)-tetrazolylacetamido]-3-methyl-Δ³-0-2-isocephem-4-carboxylate, was obtained in 22.5% yield.

Hydrogenation of the benzyl ester according to the procedure of Example34 gives the title product.

EXAMPLE 41

Replacement of the 7β-amino-3-methyl-Δ³ -0-2-isocephem-4-carboxylic acidof Examples 31-40 with an equimolar weight of 7β-amino-Δ³-0-2-isocephem-4-carboxylic acid, 7β-amino-3-benzyl-Δ³-0-2-isocephem-4-carboxylic acid or 7β-amino-3-phenethyl-Δ³-0-2-isocephem-4-carboxylic acid, respectively, produces thecorresponding 7β-acylamido-Δ³ -0-2-isocephem-4-carboxylic acids,7β-acylamido-3-benzyl-Δ³ -0-2-isocephem-4-carboxylic acids and7β-acylamido- 3-phenethyl-Δ³ -0-2-isocephem-4-carboxylic acids,respectively.

EXAMPLE 42

Repeating the general N-acylation procedures of the examples above toreact the following acylating agents with 7β-amino-3-methyl-Δ³-0-2-isocephem-4-carboxylic acid (or an ester or salt thereof), thefollowing products are obtained after removal of any functional blockinggroups.

    __________________________________________________________________________    4-nitrophenylacetyl chloride                                                                    7β-(4-nitrophenylacetamido)-                                             3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           p-fluorophenylacetyl chloride                                                                   7β-(p-fluorophenylacetamido)-                                            3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           p-acetoxyphenylacetyl chloride                                                                  7β-(p-acetoxyphenylacetamido)-                                           3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           o-chlorophenylacetyl chloride                                                                   7β-(o-chlorophenylacetamido)-                                            3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           3-thienylacetyl chloride                                                                        7β-(3-thienylacetamido)-                                                 3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           p-aminophenylacetyl chloride                                                                    7β-(p-aminophenylacetamido)-                                             3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           p-methylphenylacetyl chloride                                                                   7β-(p-methylphenylacetamido)-                                            3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           4-guanidinophenylacetyl                                                                         7β-(4-guanidinophenylacetamido)-                       chloride hydrochloride                                                                          3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic                                                4-isopropylphenylacetyl                                                                         7β-(4-isopropylphenylacetamido)-                       chloride          3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           4-methylthiophenylacetyl                                                                        7β-(4-methylthiophenylacetamido)-                      chloride          3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           4-cyanophenylacetyl chloride                                                                    7β-(4-cyanophenylacetamido)-                                             3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           4-methoxyphenylacetyl chloride                                                                  7β-(4-methoxyphenylacetamido)-                                           3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           2,6-dimethoxyphenylacetyl                                                                       7β-(2,6-dimethoxyphenylacetamido)-                     chloride          3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           3-sulfamylphenylacetyl                                                                          7β-(3-sulfamylphenylacetamido)-                        chloride          3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           2-methyl-4-chlorophenyl-                                                                        7β-(2-methyl-4-chlorophenyl-                           acetyl chloride   acetamido)-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                 sydnone-3-acetyl chloride                                                                       7β-(sydnone-3-acetamido)-                                                3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           sydnone-4-acetyl chloride                                                                       7β-(sydnone-4-acetamido)-                                                3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           2-furylacetyl chloride                                                                          7β-(2-furylacetamido)-                                                   3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           3-furylacetyl chloride                                                                          7β-(3-furylacetamido)-                                                   3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           1,2,5-thiadiazole-3-acetyl                                                                      7β-(1,2,5-thiadiazole-3-                               chloride          acetamido)-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                 1-cyclohexenylacetyl chloride                                                                   7β-(1-cyclohexenylacetamido)-                                            3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           1,4-cyclohexadienylacetyl                                                                       7-β-(1,4-cyclohexadienylacetamido)-                    chloride          3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           3-(1,4-cyclohexadien-1-yl)                                                                      7β-[3-(1,4-cyuclohexadien-1-yl)-                       propionyl chloride                                                                              propionamido]-3-methyl-Δ.sup.3 -0-2-                                    isocephem-4-carboxylic acid                                 isothiazol-4-yl-acetic acid                                                                     7β-(isothiazol-4-yl-acetamido)-                                          3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           isothiazol-5-yl-acetic acid                                                                     7β-(isothiazol-5-yl-acetamido)-                                          3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           isothiazol-3-yl-acetic acid                                                                     7β-(isothiazol-3-yl-acetamido)-                                          3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           5-phenyl-1,3,4-thiadiazolyl-2-                                                                  7β-(5-phenyl-1,3,4-thiadiazol-2-                                         yl-acetamido)-3-methyl-Δ.sup.3 -0-2-                                    isocephem-4-carboxylic acid                                 thiazol-2-yl-acetyl chloride                                                                    7β-(thiazol-2-yl-acetamido)-                                             3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           imidazol-2-yl-acetyl chloride                                                                   7β-(imidazol-2-yl-acetamido)-                                            3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           1,2,3-triazol-4-yl-acetic                                                                       7β-(1,2,3-triazol-4-yl-acetamido)-                     acid              3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           oxazol-2-yl-acetyl chloride                                                                     7β-oxazol-2-yl-acetamido)-                                               3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           4-pyridylacetyl chloride                                                                        7β-(4-pyridylacetamido)-                                                 3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           3-pyridylacetyl chloride                                                                        7β-(3-pyridylacetamido)-                                                 3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           3-phenylpropionyl chloride                                                                      7β-(3-phenylpropionamido)-                                               3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           3-(p-chlorophenyl)propionyl                                                                     7β-[3-(p-chlorophenyl)propionamido]-                   chloride          3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           3-(p-methoxyphenyl)propionyl                                                                    7β-[3-p-methoxyphenyl)propionamido]-                   chloride          3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           3-(p-sulfamylphenyl)propionyl                                                                   7β-[3-(p-sulfamylphenyl)propion-                       chloride          amido]-3-methyl-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                           3-(3,4-dimethoxyphenyl)-                                                                        7β-[3-(3,4-dimethoxyphenyl)-                           propionyl chloride                                                                              propionamido]-3-methyl-Δ.sup.3 -0-2-                                    isocephem-4-carboxylic acid                                 3-(p-hydroxyphenyl)propionic                                                                    7β-[3-(p-hydroxyphenyl)propion-                        acid              amido]-3-methyl-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                           3-(p-nitrophenyl)propionic                                                                      7β-[3-(p-nitrophenyl)propionamido]-                    acid              3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           3-(2-thienyl)propionyl                                                                          7β-[3-(2-thienyl)propionamido]-                        chloride          3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           3-(3-thienyl)propionyl                                                                          7β-[3-(3-thienyl)propionamido]-                        chloride          3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           cyclohexylacetic acid                                                                           7β-(cyclohexylacetamido)-                                                3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           3-phenyl-5-methylisoxazol-                                                                      7β-(3-phenyl-5-methylisoxazol-4-                       4-yl-acetic acid  yl-acetamido)-3-methyl-Δ.sup.3 -0-2-                                    isocephem-4-carboxylic acid                                 o-aminomethylphenylacetic                                                                       7β-(o-aminomethylphenylacetamido)-                     acid              3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           3-methoxy-4-furazanacetyl                                                                       7β-(3-methoxy-4-furazanacetamido)-                     chloride          3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                           __________________________________________________________________________

EXAMPLE 43

When the acylating agents listed in Example 42 are reacted according tothe general N-acylation procedures of the examples above with7β-amino-Δ³ -0-2-isocephem-4-carboxylic acid, 7β-amino-3-benzyl-Δ³-0-2-isocephem-4-carboxylic acid and 7β-amino-3-phenethyl-Δ³-0-2-isocephem-4-carboxylic acid, respectively, there are produced thecorresponding 7β-acylamino-Δ³ -0-2-isocephem-4-carboxylic acids,7β-acylamino-3-benzyl-Δ³ -02-isocephem-4-carboxylic acids and7β-acylamino-3-phenethyl-Δ³ -0-2-isocephem-4-carboxylic acids,respectively.

EXAMPLE 44

Repeating the general N-acylation procedures of the examples above toreact the following acylating agents with 7β-amino-3-methyl-Δ³-0-2-isocephem-4-carboxylic acid (or an ester or salt thereof), thefollowing products are obtained after removal of any functional blockinggroups.

    ______________________________________                                        p-nitrophenoxyacetyl acid                                                                    7β-(p-nitrophenoxyacetamido)-                                            3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                              p-fluorophenoxyacetyl acid                                                                   7β-(p-flourophenoxyacetamido)-                                           3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                              o-chlorophenoxyacetic acid                                                                   7β-(o-chlorophenoxyacetamido)-                                           3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                              p-sulfamylphenoxyacetic                                                                      7β-(p-sulfamylphenoxyacetamido)-                          acid           3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                              p-methylphenoxyacetic                                                                        7β-(p-methylphenoxyacetamido)-                            acid           3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                              4-hydroxyphenoxyacetic                                                                       7β-(4-hydroxyphenoxyacetamido)-                           acid           3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                              2,4-dichlorophenoxyacetic                                                                    7β-(2,4-dichlorophenoxyacetamido)-                        acid           3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                              2,6-dimethoxyphenoxyacetic                                                                   7β-(2,6-dimethoxyphenoxyaceta-                            acid           mido)-3-methyl-Δ.sup.3 -0-2-isocephem-                                  4-carboxylic acid                                              4-cyanophenoxyacetic acid                                                                    7β-(4-cyanophenoxyacetamido)-                                            3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                              α-phenoxypropionic acid                                                                7β-(α-phenoxypropionamido)-                                        3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                              α-(2-chlorophenoxy)-                                                                   7β-(α-(2-chlorophenoxy)propion-                     propionic acid amido]-3-methyl-Δ.sup.3 -0-2-isocephem-                                 4-carboxylic acid                                              α-(2,4-dichlorophenoxy)-                                                               7β-[α-(2,4-dichlorophenoxy)-                        n-butyric acid n-butyramido]-3-methyl-Δ.sup.3 -0-2-                                    isocephem-4-carboxylic acid                                    α-phenoxyphenylacetic                                                                  7β-(αphenoxyphenylacetamido)-                       acid           3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                              α-phenoxybutyric acid                                                                  7β-(α-phenoxybutyramido)-                                          3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                              4-trifluoromethylphenoxy-                                                                    7β-(4-trifluoromethylphenoxy-                             acetic acid    acetamido)-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                    benzyloxyacetyl chloride                                                                     7β-(benzyloxyacetamido)-                                                 3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                              β-naphthoxyacetyl                                                                       7β-(β-naphthoxyacetamido)-                           chloride       3-methyl-Δ.sup.3 -0-2-isocephem-                                        4-carboxylic acid                                              ______________________________________                                    

EXAMPLE 45

Replacement of the 7β-amino-3-methyl-Δ³ -0-2-isocephem-4-carboxylic acidof Example 44 with 7β-amino-Δ³ -0-2-isocephem-4-carboxylic acid,7β-amino-3-benzyl-Δ³ -0-2-isocephem-4-carboxylic acid and7β-amino-3-phenethyl-Δ³ -0-2-isocephem-4-carboxylic acid (or an ester orsalt thereof), respectively, produces the corresponding 7β-acylamino-Δ³-0-2-isocephem-4-carboxylic acids, 7β-acylamino-3-benzyl-Δ³-0-2-isocephem-4-carboxylic acids and 7β-acylamino-3-phenethyl-Δ³-0-2-isocephem-4-carboxylic acids, respectively.

EXAMPLE 46

Following the general N-acylation methods of the preceeding examples,the compounds listed below are prepared by acylating7β-amino-3-methyl-Δ³ -0-2-isocephem-4-carboxylic acid with an acylatingacid of the formula ##STR238## or a functional equivalent, e.g. acidhalide, thereof.

    ______________________________________                                         ##STR239##                                                                   R.sup.a                R.sup.b    R.sup.c                                     ______________________________________                                         ##STR240##            H          H                                            ##STR241##            H          H                                            ##STR242##            H          H                                            ##STR243##            H          H                                            ##STR244##            H          H                                            ##STR245##            H          H                                            ##STR246##            H          H                                           4-pyridyl              H          H                                           3-pyridyl              H          H                                            ##STR247##            H          H                                            ##STR248##            H          H                                           imidazolyl (2)         H          H                                           imidazolinyl (2)       H          H                                           thiazolyl (2)          H          H                                           thiazolinyl (2)        H          H                                           triazolyl (2)          H          H                                           1-methyl-              H          H                                           imidazolyl (2)                                                                2-thienyl              H          H                                           3-thienyl              H          H                                           n-butyl                H          H                                           isobutyl               H          H                                           2-acetamido-           H          H                                           thiazol-5-yl                                                                  2-phenyl-1,3,4-        H          H                                           thiadiazol-5-yl                                                               2-methyl-1,3,4-        H          H                                           oxadiazol-5-yl                                                                ______________________________________                                    

EXAMPLE 47

Replacement of the 7β-amino-3-methyl-Δ³ -0-2-isocephem-4-carboxylic acidof Example 46 with 7β-amino-Δ³ -0-2-isocephem-4-carboxylic acid,7β-amino-3-benzyl-Δ³ -0-2 isocephem-4-carboxylic acid and7β-amino-3-phenethyl-Δ³ -0-2-isocephem-4-carboxylic acid (or an ester orsalt thereof), respectively, produces the corresponding 7β-acylamino-Δ³-0-2-isocephem-4-carboxylic acids, 7β-acylamino-3-benzyl-Δ³-0-2-isocephem-4-carboxylic acids and 7β-acylamino-3-phenethyl-Δ³ -0-2-isocephem-4-carboxylic acids, respectively.

EXAMPLE 48

Following the general N-acylation methods of the proceeding examples,the compounds listed below are prepared by acylation of7β-amino-3-methyl-Δ³ -0-2-isocephem-4-carboxylic acid with theappropriate acylating acid of the general formula

    R.sup.a --COOH

or a functional equivalent thereof. ##STR249##7β-(3-phenyl-5-methyl-isoxazol-4-ylcarboxamido)-3-methyl-Δ.sup.3-0-2-isocephem-4-carboxylic acid;

7β-[3-(2,6-dichlorophenyl)-5-methylisoxazol-4-ylcarboxamido]-3-methyl-Δ³-0-2-isocephem-4-carboxylic acid;

7β-(2,6-dichlorobenzamido)-3-methyl-Δ³ -0-2-isocephem-4-carboxylic acid;

7β-(2-phenylbenzamido)-3-methyl-Δ³ -0-2-isocephem-4-carboxylic acid;

7β-(2-aminomethylbenzamido)-3-methyl-Δ³ -0-2-isocephem-4-carboxylicacid;

7β-(2-carboxybenzamido)-3-methyl-Δ³ -0-2-isocephem-4-carboxylic acid;

7β-(cyclopentanecarboxamido)-3-methyl-Δ³ -0-2-isocephem-4-carboxylicacid;

7β-(1-aminocyclohexanecarboxamido)-3-methyl-Δ³-0-2-isocephem-4-carboxylic acid;

7β-(cyclohexanecarboxamido)-3-methyl-Δ³ -0-2-isocephem-4-carboxylicacid;

7β-(1,4-cyclohexadienylcarboxamido)-3-methyl-Δ³-0-2-isocephem-4-carboxylic acid;

7β-(4-nitrobenzamido)-3-methyl-Δ³ -0-2-iosocephem-4-carboxylic acid;

7β-(4-methylbenzamido)-3-methyl-Δ³ -0-2-isocephem-4-carboxylic acid;

7β-(o-methoxybenzamido)-3-methyl-Δ³ -0-2-isocephem-4-carboxylic acid;

7β-(o-bromobenzamido)-3-methyl-Δ³ -0-2-isocephem-4-carboxylic acid;

7β-(p-ethoxybenzamido)-3-methyl-Δ³ -0-2 -isocephem-4-carboxylic acid;

7β-(o-acetamido)-3-methyl-Δ³ -0-2-isocephem-4-carboxylic acid;

7β-(p-allylbenzamido)-3-methyl-Δ³ -0-2-isocephem-4-carboxylic acid;

7β-(2,5-dihydroxybenzamido)-3-methyl-Δ³ -0-2-isocephem-4-carboxylicacid;

7β-(2-ethoxy-1-naphthamido)-3-methyl-Δ³ -0-2-isocephem-4-carboxylicacid;

7β-(2-methoxy-1-naphthamido)-3-methyl-Δ³ -0-2-isocephem-4-carboxylicacid;

7β-(o-dimethylaminobenzamido)-3-methyl-Δ³ -0-2-isocephem-4-carboxylicacid;

7β-(benzamido)-3-methyl-Δ³ -0-2-isocephem-4-carboxylic acid;

7β-(p-chlorobenzamido)-3-methyl-Δ³ -0-2-isocephem-4-carboxylic acid;

7β-(2-thienylcarboxamido)-3-methyl-Δ³ -0-2-isocephem-4-carboxylic acid;

7β-(3-thienylcarboxamido)-3-methyl-Δ³ -0-2-isocephem-4-carboxylic acid;

7β-(2-furylcarboxamido)-3-methyl-Δ³ -0-2-isocephem-4-carboxylic acid;

7β-(3-furylcarboxamido)-3-methyl-Δ³ -0-2-isocephem-4-carboxylic acid;7β-(2'-chlorocyclobutanecarboxamido)-3-methyl-Δ³-0-2-isocephem-4-carboxylic acid;

7β-(3'-fluorocyclopentanecarboxamido)-3-methyl-Δ³-0-2-isocephem-4-carboxylic acid;

7β-(3'-methylcyclopentanecarboxamido)-3-methyl-Δ³-0-2-isocephem-4-carboxylic acid;

7β-(3'-methoxycyclopentanecarboxamido)-3-methyl-Δ³-0-2-isocephem-4-carboxylic acid;

7β-(α-naphthamido)-3-methyl-Δ³ -0-2-isocephem-4-carboxylic acid;

7β-(β-naphthamido)-3-methyl-Δ³ -0-2-isocephem-4-carboxylic acid;

7β-(1-aminocyclopentanecarboxamido)-3-methyl-Δ³-0-2-isocephem-4-carboxylic acid;

7β-(1-aminocycloheptanecarboxamido)-3-methyl-Δ³-0-2-isocephem-4-carboxylic acid;

7β-(1-cyclohexenecarboxamido)-3-methyl-Δ³ -0-2-isocephem-4-carboxylicacid.

EXAMPLE 49

Replacement of the 7β-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylic acidof Example 48 with 7β-amino-Δ³ -O-2-isocephem-4-carboxylic acid,7β-amino-3-benzyl-Δ³ -O-2-isocephem-4-carboxylic acid and7β-amino-3-phenethyl-Δ³ -O-2-isocephem-4-carboxylic acid (or an ester orsalt thereof), respectively, produces the corresponding 7β-acylamino-Δ³-O-2-isocephem-4-carboxylic acids, 7β-acylamino-3-benzyl-Δ³-O-2-isocephem-4-carboxylic acids and 7β-acylamino-3-phenethyl-Δ³-O-2-isocephem-4-carboxylic acids, respectively.

EXAMPLE 50

Repeating the general N-acylation procedures of the above examples toreact trityl chloride with 7β-amino-3-methyl-Δ³-O-2-isocephem-4-carboxylic acid (or an ester or salt thereof), there isobtained after removal of any carboxyl-protecting group,7β-triphenylmethylcarboxamido-3-methyl-Δ³ -O-2-isocephem-4-carboxylicacid.

Replacement of the 7β-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylic acidin the above procedure with 7β-amino-Δ³ -O-2-isocephem-4-carboxylicacid, 7β-amino-3-benzyl-Δ³ -O-2-isocephem-4-carboxylic acid and7β-amino-3-phenethyl-Δ³ -O-2-isocephem-4-carboxylic acid, respectively,produces 7β-triphenylmethylcarboxamido-Δ³ -O-2-isocephem-4-carboxylicacid, 7β-triphenylmethylcarboxamido-3-benzyl-Δ³-O-2-isocephem-4-carboxylic acid and7β-triphenylmethylcarboxamido-3-phenethyl-Δ³ -O-2-isocephem-4-carboxylicacid, respectively.

EXAMPLE 51

Following the acylation methods of the preceding examples and inparticular those disclosed in U.S. Pat. No. 3,546,219, the compoundslisted below are prepared by reacting 7β-amino-3-methyl-Δ³-O-2-isocephem-4-carboxylic acid (or an ester or salt thereof), with theappropriate acylating agent. ##STR250##

R^(y)

phenyl

p-acetamidophenyl

p-methoxyphenyl

p-methylphenyl

2-methoxy-5-methylphenyl

m-chlorophenyl

o-nitrophenyl

2,4- dichlorophenyl

α-naphthyl

2-phenanthryl

p-aminophenyl

2-thienyl

p-dimethylaminophenyl.

EXAMPLE 52

Replacement of the 7β-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylic acidof Example 51 with 7β-amino-Δ³ -O-2-isocephem-4-carboxylic acid,7β-amino-3-benzyl-Δ³ -O-2-isocephem-4-carboxylic acid and7β-amino-3-phenethyl-Δ³ -O-2-isocephem-4-carboxylic acid (or an ester orsalt thereof), respectively, produces the corresponding 7β-acylamino-Δ³-O-2-isocephem-4-carboxylic acids, 7β-acylamino-3-benzyl-Δ³-O-2-isocephem-4-carboxylic acids and 7β-acylamino-3-phenethyl-Δ³-O-2-isocephem-4-carboxylic acids, respectively.

EXAMPLE 53

Following the acylation methods of the preceding examples and inparticular those disclosed in U.K. Pat. Nos. 1,296,081 and 1,294,541,the compounds listed below are prepared by reacting 7β-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylic acid or an ester or saltthereof with an acylating agent of the formula ##STR251## or afunctional equivalent thereof.

    ______________________________________                                         ##STR252##                                                                   R.sup.a     X     X'      R.sup.i                                             ______________________________________                                        phenyl      O     imino   NH.sub.2                                            2-thienyl   O     imino   NH.sub.2                                            3-thienyl   O     imino   NH.sub.2                                            m-nitrophenyl                                                                             O     imino   NH.sub.2                                            m-aminophenyl                                                                             O     imino   NH.sub.2                                            p-methylphenyl                                                                            O     imino   NH.sub.2                                            p-chlorophenyl                                                                            O     imino   NH.sub.2                                            p-methoxyphenyl                                                                           O     imino   NH.sub.2                                            p-hydroxyphenyl                                                                           O     imino   NH.sub.2                                            p-dimethylamino-                                                                          O     imino   NH.sub.2                                            phenyl                                                                        3,4-dimethyoxy-                                                                           O     imino   NH.sub.2                                            phenyl                                                                        m-methoxyphenyl                                                                           O     imino   NH.sub.2                                            p-acetamidophenyl                                                                         O     imino   NH.sub.2                                            m-hydroxyphenyl                                                                           O     imino   NH.sub.2                                            3,5-dichloro-4-                                                                           O     imino   NH.sub.2                                            hydroxyphenyl                                                                 3-chloro-4- O     imino   NH.sub.2                                            hydroxyphenyl                                                                 phenyl      O     O       2-furyl                                             2-thienyl   O     O       2-furyl                                             3-thienyl   O     2-furyl                                                     phenyl      O     O       phenyl                                              2-thienyl   O     O       phenyl                                              phenyl      O     O       2-thienyl                                           p-chlorophenyl                                                                            O     O       2-furyl                                             p-hydroxyphenyl                                                                           O     O       2-furyl                                             3-chloro-4- O     O       2-furyl                                             hydroxyphenyl                                                                 3,5-dichloro-4-                                                                           O     O       2-furyl                                             hydroxyphenyl                                                                 m-aminophenyl                                                                             O     O       2-furyl                                             p-methylphenyl                                                                            O     O       2-furyl                                             p-dimethylamino-                                                                          O     O       2-furyl                                             phenyl                                                                        p-methoxyphenyl                                                                           O     O       2-furyl                                             m-hydroxyphenyl                                                                           O     O       2-furyl                                             p-acetamidophenyl                                                                         O     O       2-furyl                                             m-nitrophenyl                                                                             O     O       2-furyl                                             phenyl      O     O       CH.sub.3                                            2-thienyl   O     O       CH.sub.3                                            3-thienyl   O     O       CH.sub.3                                            phenyl      O     O       CH.sub.2C.sub.6 H.sub.5                             phenyl      O     O                                                                                      ##STR253##                                         phenyl      O     O                                                                                      ##STR254##                                         phenyl      O     O                                                                                      ##STR255##                                         phenyl      O     O                                                                                      ##STR256##                                         phenyl      O     O                                                                                      ##STR257##                                         phenyl      O     O                                                                                      ##STR258##                                         phenyl      O     O                                                                                      ##STR259##                                         phenyl      O     O                                                                                      ##STR260##                                         phenyl      O     O                                                                                      ##STR261##                                         phenyl      S     O       2-furyl                                             2-thienyl   S     O       2-furyl                                             3-thienyl   S     O       2-furyl                                             p-hydroxyphyenyl                                                                          S     O       CH.sub.3                                            phenyl      O     imino   phenyl                                              phenyl      O     imino   2-thienyl                                           phenyl      O     imino   2-furyl                                             3-thineyl   O     imino   phenyl                                              phenyl      O     imino                                                                                  ##STR262##                                         ______________________________________                                    

EXAMPLE 54

Replacement of the 7β-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylic acidof Example 53 with 7β-amino-Δ³ -O-2-isocephem-4-carboxylic acid,7β-amino-3-benzyl-Δ³ -O-2-isocephem-4-carboxylic acid and7β-amino-3-phenethyl-Δ³ -O-2-isocephem-4-carboxylic acid (or an ester orsalt thereof), respectively, produces the corresponding 7β-acylamino-Δ³-O-2-isocephem-4-carboxylic acids, 7β-acylamino-3-benzyl-Δ³-O-2-isocephem-4-carboxylic acids and 7β-acylamino-3-phenethyl-Δ³-O-2-isocephem-4-carboxylic acids, respectively.

EXAMPLE 55

When 7β-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylic acid or an esteror salt thereof is acylated according to the procedures above and inparticular those disclosed in U.S. Pat. No. 3,692,779 with an acidchloride of the formula ##STR263## there are produced the compoundslisted below. ##STR264## dichloromethyl; n-propyl;

cyclopentyl;

cyclohexyl;

p-chlorobenzyl;

phenyl;

2-thienyl;

3-thienyl.

Replacement of the 7β-amino-3- methyl-Δ³ -O-2-isocephem-4-carboxylicacid in the above procedure with 7β-amino-Δ³ -O-2-isocephem-4-carboxylicacid, 7β-amino-3-benzyl-Δ³ -O-2-isocephem-4-carboxylic acid and7β-amino-3-phenethyl-Δ³ -O-2-isocephem-4-carboxylic acid (or an ester orsalt thereof), respectively, produces the corresponding 7β-acylamino-Δ³-O-2-isocephem-4-carboxylic acids, 7β-acylamino-3-benzyl-Δ³-O-2-isocephem-4-carboxylic acids and 7β-acylamino-3-phenethyl-Δ³-O-2-isocephem-4-carboxylic acids, respectively.

EXAMPLE 56

When the 7-acylamido-3-methyl-Δ³ -O-2-isocephem-4-carboxylic acidcompounds of Example 55 are hydrogenated as by the process of U.S. Pat.No. 3,692,779, there are produced the compounds listed below. ##STR265##where R^(i) is as defined in Example 55.

Replacement of the 7-acylamido-3-methyl-Δ³ -O-2-isocephem-4-carboxylicacid compounds of the above procedure with the corresponding7-acylamido-Δ³ -O-2-isocephem-4-carboxylic acid compounds,7-acylamido-3-benzyl-Δ³ -O-2-isocephem-4-carboxylic acid compounds and7-acylamido-3-phenethyl-Δ³ -O -2-isocephem-4-carboxylic acid compounds,respectively, of Example 55 produces the corresponding compounds of theformula ##STR266## where Q is hydrogen, benzyl and phenethyl,respectively.

EXAMPLE 57

When 7β-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylic acid or an esteror salt thereof is acylated according to the procedures of the aboveexamples (and in particular the procedures disclosed in U.S. Pat. No.3,646,024) with an acid chloride of the formula ##STR267## there areproduced the compounds listed below

    ______________________________________                                         ##STR268##                                                                            R.sup.a   R.sup.i                                                    ______________________________________                                                 phenyl    phenyl                                                              phenyl    2-thienyl                                                           phenyl    2-furyl                                                             2-thienyl phenyl                                                              3-thienyl 2-furyl                                                             p-hydroxyphenyl                                                                         phenyl                                                     ______________________________________                                    

Replacement of the 7β-amino-6-methyl-Δ³ -O-2-isocephem-4-carboxylic acidin the procedure above with 7β-amino-Δ³ -O-2-isocephem-4-carboxylicacid, 7β-amino-3-benzyl-Δ³ -O-2-isocephem-4-carboxylic acid and7β-amino-3-phenethyl-Δ³ -O-2-isocephem-4-carboxylic acid (or an ester orsalt thereof), respectively, produces the corresponding 7β-acylamino-Δ³-O-2-isocephem-4-carboxylic acids, 7β-acylamino-3-benzyl-Δ³-O-2-isocephem-4-carboxylic acids and 7β-acylamino-3-phenethyl-Δ³-O-2-isocephem-4-carboxylic acids, respectively.

EXAMPLE 58

When 7β-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylic acid or an esteror salt thereof is acylated according to the procedures above and inparticular according to the methods of U.S. Pat. No. 3,778,436 with anacylating agent of the formula ##STR269## or a functional equivalentthereof, there are produced the compounds listed below.

    ______________________________________                                         ##STR270##                                                                           R.sup.a     R.sup.1                                                   ______________________________________                                                3,4-dimethoxyphenyl                                                                       ethyl                                                             p-methylphenyl                                                                            2-thienyl                                                         2,4-dichlorophenyl                                                                        ethyl                                                             5-methyl-3-phenyl                                                                         ethyl                                                             isoxazol-4-yl                                                                 2-thienyl   ethyl                                                             2-furyl     2-furyl                                                           phenyl      phenyl                                                            1,4-cyclohexadien-                                                                        methyl                                                            1-yl                                                                  ______________________________________                                    

Replacement of the 7β-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylic acidin the procedure above with 7β-amino-Δ³ -O-2-isocephem-4-carboxylicacid, 7β-amino-3-benzyl-Δ³ -O-2-isocephem-4-carboxylic acid and7β-amino-3-phenethyl-Δ³ -O-2-isocephem-4-carboxylic acid (or an ester orsalt thereof), respectively, produces the corresponding 7β-acylamino-Δ³-O-2-isocephem-4-carboxylic acids, 7β-acylamino-3-benzyl-Δ³-O-2-isocephem-4-carboxylic acids and 7β-acylamino-3-phenethyl-Δ³-O-2-cephem-4-carboxylic acids, respectively.

EXAMPLE 59

When the N-acylation and de-blocking procedures of Example 2 arerepeated with the N-carbobenzoxy-D-(-)-phenylglycine used thereinreplaced by an equimolar weight of the N-t-butoxycarbonylamino acylatingacids listed below, there are produced the following compounds.

    ______________________________________                                        D-(-)-α-(3,5-dichloro-4-                                                                 7β-[D-(-)-α-amino-α-(3,5-                                    dichloro-                                                    hydroxyphenyl)-α-(t-butoxy-                                                              4-hydroxyphenyl)acetamido]-3-                                carbonylamino)acetic acid                                                                      methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                              D-(-)-α-(3-chloro-4-                                                                     7β-[D-(-)-α-amino-α-(3-chloro-              hydroxyphenyl)-2-(t-butoxy-                                                                    4-hydroxyphenyl)acetamido]-3-                                carbonylamino)acetic acid                                                                      methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                              D-(-)-α-(p-hydroxyphenyl)-                                                               7β-[D-(-)-α-amino-α-(p-hydroxy-             α-(t-butoxycarbonylamino)-                                                               phenyl)acetamido]-3-methyl-Δ.sup.3 -                   acetic acid      0-2-isocephem-4-carboxylic acid                              2-(N-t-butoxycarbonylamino)-                                                                   7β-[α-amino-α-(2,4,6-cyclohep-              2-(2,4,6-cycloheptatrien-                                                                      tatrien-1-yl)acetamido]-3-                                   1-yl)acetic accid                                                                              methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                              D-2-(t-butoxycarbonylamino)-                                                                   7β-[D-α-amino-α-(3'-hydroxy-                2-(3'-hydroxyphenyl)acetic                                                                     phenyl)acetamido]-3-methyl-Δ.sup.3 -                   acid             0-2-isocephem-4-carboxylic acid                              D-α-(t-butoxycarbonylamino)-                                                             7β-[D-α-amino-α-(4-acetamido-               α-4-acetamidophenylacetic                                                                phenyl)-acetamido]-3-methyl-Δ.sup.3 -                  acid             0-2-isocephem-4-carboxylic acid                              D-2-(t-butoxycarbonylamino)-                                                                   7β-[D-α-amino-α-(1,4-cyclo-                 2-(1,4-cyclohexadienyl)-                                                                       hexadienyl)-acetamido]-3-                                    acetic acid      methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                              D-3-(t-butoxycarbonylamino)-                                                                   7β-[D-3'-amino-3'(1,4-cyclo-                            3-(1,4-cyclohexadienyl)-                                                                       hexadienyl)propionamido]-3-                                  propionic acid   methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                              D-(-)-α-(2-thienyl)-α-                                                             7β-[D-α-amino-α-(2-thienyl)-                (t-butoxycarbonylamino)-                                                                       acetamido]-3-methyl-Δ.sup.3 -O-2-                      acetic acid      isocephem-4-carboxylic acid  54 carboxylic                                    -D-(-)-α-(3-thienyl)-α- 7β-[D-α-am                     ino-α-(3-thienyl)-                                     (t-butoxycarbonylamino)-                                                                       acetamido]-3-methyl-Δ.sup.3 -O-2-                      acetic acid      isocephem-4-carboxylic acid                                  D-α-(t-butoxycarbonylamino)-                                                             7β-[D-α-amino-α-(o-hydroxy-                 α-(o-hydroxyphenyl)acetic                                                                phenyl)acetamido]-3-methyl-Δ.sup.3 -                   acid             0-2-isocephem-4-carboxylic acid                              D-α-(t-butoxycarbonylamino)-                                                             7β-[D-α-amino-α-(p-nitrophenyl)-            α-(p-nitrophenyl)acetic                                                                  acetamido]-3-methyl-Δ.sup.3 -O-2-                      acid             isocephem-4-carboxylic acid                                  D-α-(t-butoxycarbonylamino)-                                                             7β-[-αamino-α-(p-methoxy-                                    phenyl)-                                                     α-(p-methoxyphenyl)acetic                                                                acetamido]-3-methyl-Δ.sup.3 -O-2-                      acid             isocephem-4-carboxylic acid                                  D-α-(t-butoxycarbonylamino)-                                                             7β-[D-α-amino-α-(p-cyanophenyl)-            α-(p-cyanophenyl)acetic                                                                  acetamido]-3-methyl-Δ.sup.3 -O-2-                      acid             isocephem-4-carboxylic acid                                  D-α-(t-butoxycarbonylamino)-                                                             7β-[D-α-amino-α-(p-methylthio-              α-(p-methylthiophenyl)acetic                                                             phenyl)acetamido]-3-methyl-Δ.sup.3 -                   acid             O-2-isocephem-4-carboxylic acid                              D-α-(t-butoxycarbonylamino)-                                                             7β-[D-α-amino-α -(p-isopropyl-              α-(p-isopropylphenyl)acetic                                                              phenyl)acetamido]-3-methyl-Δ.sup.3 -                   acid             O-2-isocephem-4-carboxylic acid                              D-α-(t-butoxycarbonylamino)-                                                             7β-[D-α-amino-α-(o-sulfamyl-                                 phenyl)-                                                     α-(o-sulfamylphenyl)acetic                                                               acetamido]-3-methyl-Δ.sup.3 -O-2-                      acid             isocephem-4-carboxylic acid                                  D-α-(t-butoxycarbonylamino)-                                                             7β-[D-α-amino-α-(o-amino-                   α-(o-aminomethylphenyl)-                                                                 phenyl)acetamido[-3-methyl-Δ.sup.3 -                   acetic acid      O-2-isocephem-4-carboxylic acid                              D-α-(t-butoxycarbonylamino)-                                                             7β-[D-α-amino-α-(o-dimethyl-                                 amino-                                                       α-(o-dimethylaminophenyl)-                                                               phenyl)acetamido]-3-methyl-Δ.sup.3 -                   acetic acid      0-2-isocephem-4-carboxylic acid                              D-α-(t-butoxycarbonylamino)-                                                             7β-[D-α-amino-α-(4-chloro-2-                                 thienyl)-                                                    α-(4-chloro-2-thienyl)acetic                                                             acetamido]-3-methyl-Δ.sup.3 -O-2-                      acid             isocephem-4-carboxylic acid                                  Dα-α(t-butoxycarbonylamino)-                                                       7β-[D-α-amino-α-(cyclohexyl)-               α-(cyclohexyl)acetic acid                                                                acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                  D-α-(t-butoxycarbonylamino)-                                                             7β-[D-α-amino-α-(3-trifluoro-               α-(3-trifluoromethylphenyl)-                                                             methylphenyl)acetamido]-3-                                                    methyl-                                                      acetic acid      Δ.sup.3 -0-2-isocephem-4-carboxylic                                     acid                                                         ______________________________________                                    

EXAMPLE 60

When 7β-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylic acid or an esteror salt thereof is acylated according to the general procedures of thepreceding examples with the acylating agents listed below (suitablyprotected), there are produced the following compounds.

    ______________________________________                                        α-amino-α-(1-cyclohexenyl)-                                                       7β-[α-amino-α-(1-cyclohexenyl)-              acetic acid     acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   α-amino-α-(isothiazol-4-                                                          7β-[α-amino-α-(isothiazol-4-                 yl)acetyl chloride                                                                            yl)acetamido]-3-methylΔ.sup.3 -0-2-                                     isocephem-4-carboxylic acid                                   α-amino-α-(5-phenyl-1,3,4-                                                        7β-[α-amino-α-(5-phenyl-1,3,4-               thiadiazol-2-yl)acetyl                                                                        thiadiazol-2-yl)acetamido]-3-                                 chloride        methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                               α-amino-α-(5-phenyl-1,3,4-                                                        7β-[α-amino-α-(5-phenyl-1,3,4-               oxadiazol-2-yl)acetyl                                                                         oxadiazol-2-yl)acetamido]-3-                                  chloride        methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                               α-amino-α-(3-methyl-1,2,5-                                                        7β-[α-amino-α-(3-methyl-1,2,5-               oxadiazol-4-yl)acetyl                                                                         oxadiazol-4-yl)acetamido]-3-                                  chloride        methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                               α-amino-α-(oxazol-2-yl)-                                                          7β-[α-amino-α-(oxazol-2-yl)-                 acetyl chloride acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   α-amino-α-(1H)-tetrazolyl-                                                        7β-[α-amino-α-(1H)-tetrazolyl-               acetyl chloride acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   α-amino-α-4-isoxazolyl-                                                           7β-[α-amino-α-isoxazolyl)-                   acetyl chloride acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   α-amino-α-(2-thiazolyl)-                                                          7β-(α-amino-α-(2-thiazolyl)-                 acetyl chloride acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   α-amino-α-(2-furyl)-                                                              7β-[α-amino-α-(2-furyl)-                     acetyl chloride acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   α-amino-α-(1,2,5-thiadiazol-                                                      7β-[α-amino-α-(1,2,5-thiadiazol-             3-yl)acetyl chloride                                                                          3-yl)acetamido]-3-methyl-Δ.sup.3 -0-2-                                  isocephem-4-carboxylic acid                                   α-amino-α-(3-furyl)acetyl                                                         7β-[α-amino-α-(3-furyl)-                     chloride        acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   ______________________________________                                    

EXAMPLE 61

When the 7β-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylic acid in theprocedures of Examples 59 and 60 is replaced by 7β-amino-Δ³-O-2-isocephem-4-carboxylic acid, 7β-amino-3-benzyl-Δ³-O-2-isocephem-4-carboxylic acid and 7β-amino-3-phenethyl-Δ³-O-2-isocephem-4-carboxylic acid, respectively, there are produced thecorresponding 7-acylamino-Δ³ -O-2-isocephem-4-carboxylicacids,7-acylamino-3-benzyl-Δ³ -O-2-isocephem-4-carboxylic acids and7-aceylamino-3-phenethyl-Δ³ -O-2-isocephem-4-carboxylic acids,respectively.

EXAMPLE 62

When benzyl 7β-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylate isacylated according to the procedure of Example 27 with the acylatingagents listed below (suitably protected if necessary), there areproduced the following compounds after removal of any protecting groups:

    ______________________________________                                        D-3-chloromandelic acid                                                                       7β-[D-α-hydroxy-α-(3-chloro-                                 phenyl)-                                                                      acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   D-2-trifluoromethyl-                                                                          7β-[D-α-hydroxy-α-(2-trifluoro-              mandelic acid   methylphenyl)acetamido]-3-methyl-                                             Δ.sup.3 -0-2-isocephem-4-carboxylic acid                D-3-nitromandelic acid                                                                        7β-[D-α-hydroxy-α-(3-nitrophenyl)-                           acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   D-p-hydroxymandelic acid                                                                      7β-[D-α-hydroxy-α-(p-hydroxy-                                phenyl)                                                                       acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   D-3-chloro-4-hydroxyman-                                                                      7β-[D-α-hydroxy-α-(3-chloro-4-               delic acid      hydroxyphenyl)acetamido]-3-methyl-                                            Δ.sup.3 -0-2-isocephem-4-carboxylic acid                D-3,5-dichloro-4-hydroxy-                                                                     7β-[D-α-hydroxy-α-(3,5-dichloro-             mandelic acid   4-hydroxyphenyl)acetamido]-3-                                                 methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                               D-o-methylaminomandelic                                                                       7β-[D-α-hydroxy-α-(o-methylamino-            acid            phenyl)acetamido]-3-methyl-Δ.sup.3 -0-2-                                isocephem-4-carboxylic acid                                   D-p-methoxymandelic acid                                                                      7β-[D-α-hydroxy-α-(p-methoxy-                                phenyl)acetamido]-3-methyl-Δ.sup.3 -0-2-                                isocephem-4-carboxylic acid                                   D-m-methylthiomandelic                                                                        7β-[D-α-hydroxy-α-(m-methylthio-             acid            phenyl)acetamido]-3-methyl-Δ.sup.3 -0-2-                                isocephem-4-carboxylic acid                                   D-m-iodomandelic acid                                                                         7β-[D-α-hydroxy-α-(m-iodophenyl)-                            acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   4-isoxazoleglycolic acid                                                                      7β-[α-hydroxy-α-(4-isoxazolyl)-                              acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   4-thiazoleglycolic acid                                                                       7β-[α-hydroxy-α-(4-thiazolyl)-                               acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   4-oxazoleglycolic acid                                                                        7β-[α -hydroxy-α-(4-oxazolyl)-                               acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   3-isothiazoleglycolic acid                                                                    7β-[α-hydroxy-α-(3-isothiazolyl)-                            acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   1,2,3-triazole-4-glycolic                                                                     7β-[α-hydroxy-α-(1,2,3-triazol-              acid            4-yl)acetamido]-3-methyl-Δ.sup.3 -0-2-                                  isocephem-4-carboxylic acid                                   5-isoxazoleglycolic acid                                                                      7β-[α-hydroxy-α-(5-isoxazolyl)-                              acetamido-3-methyl-Δ.sup.3 -0-2-                                        isocephem-4-carboxylic acid                                   1,2,4-triazole-3-glycolic                                                                     7β-[α-hydroxy-α-(1,2,4-triazol-              acid            3-yl)acetamido]-3-methyl-Δ.sup.3 -0-2-                                  isocephem-4-carboxylic acid                                   2-thienylglycolic acid                                                                        7β-[α-hydroxy-α-(2-thienyl)-                                 acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   3-thienylglycolic acid                                                                        7β-[α-hydroxy-α-(3-thienyl)-                                 acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   1,4-cyclohexadien-1-yl-                                                                       7β-[α-hydroxy-α-(1,4-cyclo-                                  hexadien-                                                     glycolic acid   1-yl)acetamido]-3-methyl-Δ.sup.3 -0-2-                                  isocephem-4-carboxylic acid                                   1-cyclohexenylglycolic                                                                        7β-[α-hydroxy-α-(1-cyclohexenyl)-            acid            acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   2-pyrrolylglycolic acid                                                                       7β-[α-hydroxy-α-(2-pyrrolyl)-                                acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   2-furylglycolic acid                                                                          7β-[α-hydroxy-α-(2-furyl)-                                   acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   3-pyridylglycolic acid                                                                        7β-[α-hydroxy-α-(3-pyridyl)-                                 acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   ______________________________________                                    

EXAMPLE 62

When the benzyl 7β-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylate in theprocedure of Example 62 is replaced by benzyl 7β-amino-Δ³-O-2-isocephem-4-carboxylate, benzyl 7β-amino-3-benzyl-Δ³-O-2-isocephem-4-carboxylate and benzyl 7β-amino-3-phenethyl-Δ³-O-2-isocephem-4-carboxylate, respectively, there are produced thecorresponding 7-acylamido-Δ³ -O-2-isocephem-4-carboxylic acids,7-acylamido-3-benzyl-Δ³ -O-2-isocephem-4-carboxylic acids and7-acylamido-3-phenethyl-Δ³ -O-2-isocephem-4-carboxylic acids,respectively.

EXAMPLE 64

When benzyl 7β-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylate isacylated according to the procedure of Example 34 with the acylatingagents listed below (suitably protected if desired), there are producedthe following compounds after removal of any protecting groups:

    ______________________________________                                        p-hydroxyphenylmalonic acid                                                                   7β-[α-carboxy-α-(p-hydrophenyl)-                             acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   3-chloro-4-hydroxyphenyl-                                                                     7β-[α-carboxy-α-(3-chloro-4-                 malonic acid    hydroxyphenyl)acetamido]-3-methyl-                                            Δ.sup.3 -0-2-isocephem-4-carboxylic acid                3,5-dichloro-4-hydroxy-                                                                       7β-[α-carboxy-α-(3,5-dichloro-               phenylmalonic acid                                                                            4-hydroxyphenyl)acetamido]-3-                                                 methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                               o-chlorophenylmalonic                                                                         7β-[α-carboxy-α-(o-chlorophenyl)-            acid            acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   p-nitrophenylmalonic                                                                          7β-[α-carboxy-α-(p-nitrophenyl)-             acid            acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   p-acetoxyphenylmalonic                                                                        7β-[α-carboxy-α-(p-acetoxyphenyl)-           acid            acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   p-methoxyphenylmalonic                                                                        7β-[α-carboxy-α-(p-methoxyphenyl)-           acid            acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   p-methylthiophenylmalonic                                                                     7β-[α-carboxy-α-(p-methylthio-               acid            phenyl)acetamido]-3-methyl-Δ.sup.3 -0-2-                                isocephem-4-carboxylic acid                                   p-cyanophenylmalonic                                                                          7β-[α-carboxy-α-(p-cyanophenyl)-             acid            acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   m-isopropylphenylmalonic                                                                      7β-[α-carboxy-α-(m-isopropylphenyl)-         acid            acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   o-aminomethylphenylmalonic                                                                    7β-[α-carboxy-α-(o-aminomethyl-              acid            phenyl)acetamido]-3-methyl-Δ.sup.3 -0-2-                                isocephem-4-carboxylic acid                                   o-dimethylaminophenyl                                                                         7β-[α-carboxy-α-(o-dimethylamino-            malonic acid    phenyl)acetamido]-3-methyl-Δ.sup.3 -0-2-                                isocephem-4-carboxylic acid                                   2-thienylmalonic acid                                                                         7β-[α-carboxy-α-(2-thienyl)-                                 acetamido]-3-methyl-Δ  .sup.3 -0-2-                                     isocephem-4-carboxylic acid                                   3-thienylmalonic acid                                                                         7β-[α-carboxy-α-(3-thienyl)-                                 acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   1,4-cyclohexadienylmalonic                                                                    7β-[α-carboxy-α-(1,4-cyclo-                  acid            hexadien-1-yl)acetamido]-3-                                                   methyl-Δ.sup.3 -0-2-isocephem-4-                                        carboxylic acid                                               1-cyclohexenylmalonic                                                                         7β-[α-carboxy-α-(1-cyclo-                    acid            hexenyl)acetamido]-3-methyl-                                                  Δ.sup.3 -0-2-isocephem-4-carboxylic                                     acid                                                          2-furylmalonic acid                                                                           7β-[α-carboxy-α-(2-furyl)-                                   acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   4-pyridylmalonic acid                                                                         7β-[α-carboxy-α-(4-pyridyl)-                                 acetamido]-3-methyl-Δ.sup.3 -0-2-                                       isocephem-4-carboxylic acid                                   ______________________________________                                    

EXAMPLE 65

When benzyl 7β-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylate in theprocedure of Example 64 is replaced by benzyl 7β-amino-Δ³-O-2-isocephem-4-carboxylate, benzyl 7β-amino-3-benzyl-Δ³-O-2-isocephem-4-carboxylate and benzyl 7β-amino-3-phenethyl-Δ³-O-2-isocephem-4-carboxylate, respectively, there are produced thecorresponding 7-acylamido-Δ³ -O-2-isocephem-4-carboxylic acids,7-acylamido-3-benzyl-Δ³ -O-2-isocephem-4-carboxylic acids and7-acylamido-3-phenethyl-Δ³ -O-2-isocephem-4-carboxylic acids,respectively.

EXAMPLE 66

When 7β-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylic acid or an esteror salt thereof is acylated according to the general procedures of thepreceeding examples with an acylating agent of the formula ##STR271## ora functional equivalent thereof, there are produced the compounds listedbelow

    ______________________________________                                         ##STR272##                                                                   R.sup.a             Y                                                         ______________________________________                                        phenyl             guanidino                                                  2-thienyl          guanidino                                                  3-thienyl          guanidino                                                  1,4-cyclohexadienyl                                                                              guanidino                                                  1-cyclohexenyl     guanidino                                                  p-hydroxyphenyl    guanidino                                                  3-chloro-4-hydroxyphenyl                                                                         guanidino                                                  3,5-dichloro-4-hydroxphenyl                                                                      guanidino                                                  phenyl             ureido                                                     2-thienyl          ureido                                                     3-thienyl          ureido                                                     1-cyclohexenyl     ureido                                                     1,4-cyclohexadienyl                                                                              ureido                                                     p-hydroxyphenyl    ureido                                                     3,5-dichloro-4-hydroxyphenyl                                                                     ureido                                                     o-aminomethylphenyl                                                                              ureido                                                     p-methylphenyl     ureido                                                     m-chlorophenyl     ureido                                                     phenyl             thioureido                                                 2-thienyl          thioureido                                                 3-thienyl          thioureido                                                 1-cyclohexenyl     thioureido                                                 1,4-cyclohexadienyl                                                                              thioureido                                                 p-hydroxyphenyl    thioureido                                                 3-chloro-4-hydroxyphenyl                                                                         thioureido                                                 3,5-dichloro-4-hydroxyphenyl                                                                     thioureido                                                 o-aminomethylphenyl                                                                              methylthioureido                                           m-chlorophenyl     allylthioureido                                            phenyl             chloro                                                     2-thienyl          bromo                                                      3-thienyl          chloro                                                     1,4-cyclohexadienyl                                                                              methoxy                                                    1-cyclohexenyl     ethoxy                                                     phenyl             phenyl                                                     2-thienyl          methoxy                                                    3-thienyl          ethoxy                                                     p-hydroxyphenyl    iodo                                                       p-trifluoromethylphenyl                                                                          methoxy                                                    3,4-dichlorophenyl methoxy                                                    phenyl             acetoxy                                                    2-thienyl          acetoxy                                                    2-furyl            acetoxy                                                    p-nitrophenyl      acetoxy                                                    p-methoxyphenyl    acetoxy                                                    phenyl             propionyloxy                                               phenyl             cyano                                                      phenyl             SO.sub.3 H                                                 phenyl             azido                                                      phenyl             methylsulfonyl                                             phenyl             5-indanyloxycarbonyl                                       p-hydroxyphenyl    5-indanyloxycarbonyl                                       3-chloro-4-hydroxyphenyl                                                                         5-indanyloxycarbonyl                                       3,5-dichloro-4-hydroxyphenyl                                                                     5-indanyloxycarbonyl                                       ______________________________________                                    

EXAMPLE 67

When the 7β-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylic acid in theprocedure of Example 66 is replaced by 7β-amino-Δ³-O-2-isocephem-4-carboxylic acid, 7β-amino-3-benzyl-Δ³-O-2-isocephem-4-carboxylic acid and 7β-amino-3-phenethyl-Δ³-O-2-isocephem-4-carboxylic acid (or an ester or salt thereof),respectively, there are produced the corresponding 7β-acylamino-Δ³-O-2-isocephem-4-carboxylic acids, 7β-acylamino-3-benzyl-Δ³-O-2-isocephem-4-carboxylic acids and 7β-acylamino-3-phenethyl-Δ³-O-2-isocephem-4-carboxylic acids, respectively.

EXAMPLE 68

When the α-amino products of Examples 2, 13, 21-23 and 59-61 are reactedwith acetone according to the procedure of U.S. Pat. No. 3,303,193,there are obtained the corresponding 0-2-isocephem derivatives of theformula ##STR273## where R^(a) is as defined in the above-mentionedexamples and Q is hydrogen, methyl, benzyl or phenethyl, orpharmaceutically acceptable salts thereof.

EXAMPLE 69

When the α-amino products of Examples 2, 13, 21, 22, 23 or 59-61 arereacted with dicyanogen or cyanogen bromide or cyanogen chlorideaccording to the procedure disclosed in U.S. Pat. No. 3,796,709, thecorresponding α-cyanamino products are obtained.

EXAMPLE 70

When the α-amino products of Examples 2, 13, 21-23 or 59-61 are reactedwith a triethylamine -SO₃ complex according to the procedure of U.S.Pat. No. 3,381,001, the corresponding α-sulfoamino products areobtained.

EXAMPLE 71

When the α-amino products of Examples 2, 13, 21-23 and 59-61 are reactedwith 1-methyl-1-nitrosobiuret or a1-methyl-5-(lower)alkyl-1-nitrosobiuret according to the procedure ofU.S. Pat. No. 3,483,188, there are produced the compounds of the generalformula ##STR274## where R² is hydrogen or (lower)alkyl, Q is hydrogen,methyl, benzyl or phenethyl and R^(a) is as defined in theabovementioned examples.

EXAMPLE 72

When the 7-amino intermediates in the acylation procedures of Examples1-2, 5-8, 12 and 21-67 are replaced by the correspondingpivaloyloxymethyl, acetoxymethyl, methoxymethyl, acetonyl and phenacylesters, respectively, and the ester group of the 7-acylamido product isnot removed, there are obtained the corresponding pivaloyloxymethyl,acetoxymethyl, methoxymethyl, acetonyl and phenacyl esters,respectively, of the 7-acylamido end products.

EXAMPLE 73 Preparation of7β-[α-(2-Aminomethyl-1,4-cyclohexadienyl)acetamido]-3-methyl-.DELTA.³-O-2-isocephem-4-carboxylic acid ##STR275## A.α-(2-Aminomethyl-1,4-cyclohexadienyl)acetic acid

A solution of 16.5 g. (0.1 mole) of o-aminomethylphenylacetic acid in1.5 l of liquid ammonia (which had been treated with 50 mg. of Li toremove a trace of moisture) was slowly diluted with 500 ml. of dryt-BuOH. To the solution was added in small portions 3.4 g. (0.5 atom) ofLi over a period of 4 hours andthe mixture was stirred for 16 hours atroom temperature removing the liquid ammonia in a hood and finallyevaporated to dryness below 40° C. The residue was dissolved in 500 ml.of water and the solution was chromatographed on a column of IR-120(H³⁰, 700 ml.) resin and eluted with 1% NH₄ OH solution. Ninhydrinpositive fractions of the eluate were combined and evaporated todryness. The residue was washed with four 50 ml. portions of hot acetoneand recrystallized from 500 ml. of ethanol-water (1:1) to give 11.2 g.(67%) of α-(2-aminomethyl-1,4 -cyclohexadienyl)acetic acid as colorlessneedles. M.p. 183° C.

IR: ν_(max) ^(nuj) 1630, 1520, 1380, 1356 cm⁻¹

NMR: δD₂ O + K₂ CO₃ 2.72 (4H, s, ##STR276## 3.01 (2H, s, CH₂ CO), 3.20(2H, s, CH₂ -N), 5.78 (2H, s, ##STR277##

Anal. Calcd. for C₉ H₁₃ NO₂ : C, 64.65; H, 7.84; N, 8.38. Found: C,64.77; H, 8.06; N, 8.44.

B. α-[2-(t-Butoxycarbonylaminomethyl)-1,4-cyclohexadienyl]acetic acid

to a stirred solution of 8.0 g. (0.048 mole) ofα-(2-aminomethyl-1,4-cyclohexadienyl)acetic acid and 3.8 g. (0.096 mole)of NaOH in 150 ml. of water was added a solution of 10.3 g. (0.072 mole)of t-butoxycarbonylazide in 80 ml. of THF and the mixture was stirredfor 18 hours at room temperature. The THF was removed under reducedpressure and the residual solution was washed with ether (2 × 100 ml.),acidified with 6 N HCl and extracted with ether (3 × 100 ml.). Thecombined extracts were washed with water (2 × 100 ml.) and a saturatedNaCl solution (100 ml.), dried with Na₂ SO₄ and evaporated to dryness.The oily residue was triturated with n-hexane to give 10.5 g. (82%) ofcolorless powder melting at 113° C.

IR: ν_(max) ^(nuj) 3370, 1715, 1640, 1530, 1280, 1160 cm⁻¹.

NMR: δ_(ppm) ^(CDCl) 3 1.45 (9H, s, t-Bu-H), 2.73 (4H, s, ##STR278##3.16 (2H, s, CH₂ CO); 3.76 (2H, d, 6Hz, CH₂ N) 4.90 (1H, m, NH), 5.66(2H, s, ##STR279## 10.6 (1H, br-s, COOH).

Anal. Calcd. for C₁₄ H₂₁ NO₄ : C, 62.90; H, 7.92; N, 5.24. Found: C,63.13; H, 8.21; N, 5.26.

7β-[α-(2-t-Butoxycarbonylaminomethyl-1,4-cyclohexadienyl)acetamido]-3-methyl-Δ³-O-2-isocephem-4-carboxylic acid

To a stirred solution of equimolar amounts ofα-[2-(t-butoxycarbonylaminomethyl)-1,4-cyclohexadienyl)acetic acid and2,4-dinitrophenol in ethyl acetate is added an equimolar amount ofN,N'-dicyclohexylcarbodiimide. The reaction mixture is stirred at roomtemperature for 3 hours. The separated dicyclohexylurea is filtered off.The filtrate is evaporated to dryness to give the activated ester whichis dissolved in tetrahydrofuran. To this solution is added a solution of7β-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylic acid and triethylaminein approximately a 1:2 molar proportion, respectively, relative to theα-[2-(t-butoxycarbonylaminomethyl)-1,4-cyclohexadienyl]acetic acid. Themixture is stirred at room temperature for several hours andconcentrated in vacuo. The concentrate is washed with ether, acidifiedwith dilute mineral acid and extracted with ethyl acetate. The extractsare washed with water and saturated NaCl solution and dried to give thetitle product.

D. 7β-[α-(2-Aminomethyl-1,4-cyclohexadienyl)acetamido]-3-methyl-.DELTA.³-O-2-isocephem-4-carboxylic acid

A solution of7β-[α-(2-t-butoxycarbonylaminomethyl-1,4-cyclohexadienyl)acetamido]-3-methyl-Δ³-O-2-isocephem-4-carboxylic acid in trifluoroacetic acid is stirred at0° C. for 1 hour. To the solution is added dry ether until a precipitateforms. The precipitate is collected by filtration, suspended in waterand adjusted to pH6 to give the title product.

EXAMPLE 747β-[α-(2-Aminomethyl-1-cyclohexenyl)acetamido]-3-methyl-Δ.sup.3--O-2-isocephem-4-carboxylic acid ##STR280## A.[2-(N-t-Butoxycarbonylaminomethyl)-1-cyclohexen-1-yl]acetic acid

A solution ofα-[2-(t-butoxycarbonylaminomethyl)-1,4-cyclohexadienyl]-acetic acid(1.33 g., 5 mmoles) in 3% ammonium hydroxide (10 ml.) was hydrogenatedat 40 psi with palladium on charcoal (10%, 0.2 g.). A theoretical amountof hydrogen was taken up in 3 hours. The catalyst was removed and thefiltrate was acidified to pH 2 with dil. HCl and extracted with ethylacetate (2 × 50 ml.). The combined extracts were washed with water (20ml.), dried with Na₂ SO₄ and evaporated under reduced pressure to affordan oil (1.34 g.) which solidified on standing for several days.Recrystallization from n-hexane - ethyl acetate gave 1.2 g. titleproduct as colorless prisms melting at 118°-119° C.

IR: ν_(max) ^(nujol) 3450, 1730, 1660, 1510 cm⁻¹.

NMR: δ_(ppm) ^(CDCl) 3 1.58 (9H, s, t-butyl-H), 1.50 - 1.90 (4H, m,--CH₂ --), 1.90 - 2.20 (4H, m, allylic methylene-H), 3.18 (2H, s, CH₂--CO), 3.78 (2H, d, 6Hz, CH₂ -N), 5.00 (1H, br-s, NH), 8.98 (1H, br-s,COOH).

Anal. Calcd. for C₁₄ H₂₃ NO₄ : C, 62.43; H, 8.61; N, 5.20. Found: C,62.12; H, 8.77; N, 5.37.

B.7β-[α-(2-t-Butoxycarbonylaminomethyl-1-cyclohexenyl)acetamido]-3-methyl-Δ³-O-2-isocephem-4-carboxylic acid

To a stirred solution of equimolar amounts of[2-(N-t-butoxycarbonylaminomethyl)-1-cyclohexen-1-yl]acetic acid and2,4-dinitrophenol and ethyl acetate is added an equimolar amount ofN,N'-dicyclohexylcarbodiimide. The reaction mixture is stirred for 1hour at room temperature and the precipitated dicyclohexylurea isfiltered off. The filtrate is cooled to 5° C. and poured into a coldsolution of 7β-amino-3-methyl-Δ³ -O-2-isocephem-4-carboxylic acid andexcess triethylamine in 50% aqueous THF. The mixture is stirredovernight at room temperature and washed with ether. The aqueous layeris acidified with dilute HCl to precipitate the title product.

C. 7β-[α-(2-Aminomethyl-1-cyclohexenyl)acetamido]-3-methyl-Δ³-0-2-isocephem-4-carboxylic acid

A solution of7β-[α-(2-t-butoxycarbonylaminomethyl-1-cyclohexenyl)acetamido]-3-methyl-Δ³-0-2-isocephem-4-carboxylic acid in trifluoroacetic acid is stirred at0° C. for 1.5 hours. The mixture is diluted with ether to separate thetrifluoroacetate salt which is dissolved in water and neutralized togive the title product.

EXAMPLE 75

When the procedure of Example 12 is repeated with the benzyl7β-phenoxymethylacetamido-3-methyl-Δ³ -O-2-isocephem-4-carboxylatereplaced by an equimolar weight of the benzyl ester of each of the7-acylamido-3-methyl-Δ³ -O-2-isocephem-4-carboxylic acid products ofExamples 1-2, 13, 24-29, 31-40, 42, 44, 46, 48, 50, 51, 53, 55, 56, 57,58-60, 62, 64, 66 and 68-74, there are obtained the corresponding7-acylamido-3-carboxymethylene-Δ³ -O-2-isocephem-4-carboxylic acids (andesters) and 7-acylamido-3-carbomethoxymethylene-Δ³-O-2-isocephem-4-carboxylic acids (and esters).

EXAMPLE 76

When the 7-acylamido-3-carboxymethylene-Δ³ -O-2-isocephem-4-carboxylicacids and 7-acylamido-3-carboxymethylene-Δ³ -O-2-isocephem-4-carboxylicacids of Examples 12 and 75 are reacted with chloromethyl pivalate,chloromethyl acetate, chloromethyl methyl ether, chloroacetone andphenacyl bromide, respectively, according to the general proceduredisclosed in U.K. Pat. No. 1,229,453, there are obtained thecorresponding pivaloyloxymethyl, acetoxymethyl methoxymethyl, acetonyland phenacyl esters, respectively.

    ______________________________________                                        M.I.C. (mcg./ml.)                                                             ______________________________________                                                                  Sal.                                                Compound                                                                              S. aureus E. coli enteritidis                                                                           D. pneumoniae                               of Ex. No.                                                                            A9537     A15119  A9531   A9585                                       ______________________________________                                        24      0.6       16      >2.5    0.08                                        25      0.6       63      8       .3                                          26      >125      >125    32      32                                          27      2         16      8       1                                           28      4         >125    125     0.5                                         29      125       >125    >125    125                                         31      32        >125    >125    16                                          32      1         125     4       0.5                                         33      1         32      2       0.25                                        34      16        125     32      4                                           35      0.5       >125    >125    0.25                                        36      4         125     125     0.5                                         37      >125      >125    >125    125                                         38      125       >125    >125    32                                          39      >8        >125    >125    8                                           ______________________________________                                    

We claim:
 1. A compound having the formula ##STR281## wherein R is anacyl group of the formula

    R.sup.a C.sub.n H.sub.2n CO--                              (i)

wherein R^(a) is (a) aryl selected from the group consisting of phenyl,2-thienyl, 3-thienyl, furyl, 4-isoxazolyl, pyridyl, tetrazolyl,sydnone-3 or -4, imidazolyl, naphthoyl, quinoxalinyl, triazolyl,isothiazolyl, thiadiazolyl, thiazolyl, oxazolyl, oxadiazolyl, pyrazolyl,furazan, pyrazinyl, pyrimidinyl, pyridazinyl and triazinyl; (b)substituted aryl in which the aryl groups defined above under (a) aresubstituted by one or more chloro, bromo, iodo, fluoro, nitro, amino,cyano, (lower)alkanoyloxy, (lower)alkanoyl, (lower)alkoxyamino,(lower)alkoxy, (lower)alkyl, (lower)alkylamino, hydroxy, guanidino,(lower)alkylthio, carboxy, phenyl, halophenyl, trifluoromethyl,di(lower)alkylamino, sulfamyl, (lower)alkanoylamino,phenyl(lower)alkylamido, cycloalkylamino, allylamido,morpholinocarbonyl, pyrrolidinocarbonyl, piperidinocarbonyl,tetrahydropyridino, furfurylamido or N-alkyl-N-anilino radicals; (c) C₃-C₁₂ cycloalkyl; (d) substituted C₃ -C₁₂ cycloalkyl in which thesubstituents are one or more chloro, bromo, fluoro, iodo, nitro,trifluoromethyl, C₁ -C₄ alkyl, C₁ -C₄ alkylamino, C₁ -C₂ alkoxy or aminoradicals; (e) C₃ -C₁₂ cycloalkenyl, said cycloalkenyl group having 1 or2 double bonds; or (f) substituted C₃ -C₁₂ cycloalkenyl, saidcycloalkenyl group having 1 or 2 double bonds and being substituted byone or more chloro, bromo, fluoro, iodo, nitro, trifluoromethyl, C₁ -C₄alkyl, C₁ -C₄ alkylamino, C₁ -C₂ alkoxy or amino radicals; and n is aninteger from 1-4;

    C.sub.n H.sub.2n+1 CO--                                    (ii)

wherein n is an integer from 1-7, the alkyl portion of said acyl groupbeing straight or branched and optionally interrupted by an oxygen orsulfur atom;

    C.sub.n H.sub.2n-1 CO--                                    (iii)

wherein n is an integer from 2-7, the alkenyl portion of said acyl groupbeing straight or branched and optionally interrupted by an oxygen orsulfur atom; ##STR282## wherein R^(a) is as defined above under (i) andin addition may be benzyl, C₁ -C₆ alkyl or (lower)alkoxy carbonyl andR^(b) and R^(c) which may be the same or different each representhydrogen, phenyl, benzyl, phenethyl or C₁ -C₆ alkyl; ##STR283## whereinR^(a) is as defined bove under (i) and in addition may be benzyl or C₁-C₆ alkyl and R^(b) and R^(c) are as defined under (iv);

    R.sup.a X(CH.sub.2).sub.m CO--                             (vi)

wherein R^(a) is as defined under (i) and in addition may be benzyl; Xis oxygen or sulfur; and m is an integer of 2-5;

    R.sup.a CO--                                               (vii)

wherein R^(a) is as defined under (i); ##STR284## wherein R^(a) is asdefined under (i) and Y is hydrazino, guanidino, ureido; substitutedureido of the formula ##STR285## in which R^(p) is hydrogen or C₁ -C₈alkyl and R^(q) is hydrogen, C₁ -C₈ alkyl, C₂ -C₈ alkenyl, phenyl,benzoyl, C₁ -C₈ alkoxy-C₁ -C₈ alkyl or (carbo-C₁ -C₈ alkoxy)C₁ -C₈alkyl; allophanamido; 3-guanyl-1-ureido; 3-(2-furoyl)ureido;3-(benzoyl)ureido; cyano; cyanamino; azido; amino; a group obtained byreacting the amino group Y with acetone, formaldehyde,acetaldehyde,butyraldehyde, acetylacetone, methyl acetoacetate,benzaldehyde, salicylaldehyde, methyl ethyl ketone or ethylacetoacetate; hydroxy; (lower)alkoxy; carboxy; 5-indanyloxycarbonyl;triazolyl; tetrazolyl; halogeno; formyloxy; (lower)alkanoyloxy; sulfo;or sulfoamino; ##STR286## wherein R^(d), R³ and R^(f) which may be thesame or different may each represent C₁ -C₆ alkyl, phenyl or phenylsubstituted by one or more chloro, bromo, iodo, fluoro, trifluoromethyl,nitro, amino, cyano, (lower)alkanoyloxy, (lower)alkanoyl,(lower)alkoxyamino, (lower)alkoxy, (lower)alkyl, (lower)alkylamino,hydroxy, (lower)alkylthio, carboxy, di(lower)alkylamino or sulfamylradicals; ##STR287## wherein R^(a) is as defined under (i) and inaddition may be hydrogen, C.sub. -C₆ alkyl, halogen-substituted C₁ -C₆alkyl, phenethyl, phenoxymethyl, benzyl or R^(a) --CO-- and X is oxygenor sulfur; ##STR288## wherein Y is as defined under (viii) and n is aninteger of 1-4;

    R.sup.g CH(NH.sub.2)(CH.sub.2).sub.n CO--                  (xii)

wherein n is an integer of 1-10, or

    H.sub.2 N--C.sub.n H.sub.2n Ar(CH.sub.2).sub.m CO--

wherein m is 0 or an integer from 1-10, and n is 0, 1 or 2; R^(g) ishydrogen, (lower)alkyl, phenyl, benzyl or carboxy and Ar is p-phenyleneor 1,4-naphthylene;

    R.sup.h CO.CO--                                            (xiii)

wherein R^(h) is 2-thienyl; 3-thienyl; α-naphthyl; 2-phenanthryl or amono-, di- or tri-substituted phenyl group, the substituents beingchloro, bromo, iodo, fluoro, amino, di(lower)alkylamino, (lower)alkyl,(lower)alkoxy, nitro or (lower)alkanoylamino; ##STR289## wherein R^(a)is as defined under (i); X is oxygen or sulfur; X' is oxygen or imino;and R^(i) is (lower)alyl, cycloalkyl having 4, 5, 6 or 7 carbon atoms,monohalo (lower) alkyl, dichloromethyl, trichloromethyl, (lower)alkenylof 2-6 carbon atoms, ##STR290## n is an integer from 0 to 3 inclusiveand each of R^(k) and R^(j) is hydrogen, nitro, di(lower)alkylamino,(lower)alkanoylamino, (lower)alkanoyloxy, C₁ --C₆ alkyl, C₁ -C₆ alkoxy,sulfamyl, chloro, iodo, bromo, fluoro, or trifluoromethyl; ##STR291##wherein R^(a) is as defined under (i) and R^(i) is as defined under(xiv); or ##STR292## wherein R^(a) is as defined under (i) and R¹ is(lower)alkyl, cycloalkyl of 3-12 carbon atoms, phenyl, a monocyclicheterocyclic radical having 5 or 6 atoms exclusive of hydrogen which areC, S, N or O, no more than 2 atoms being other than C, or a substitutedmonocyclic heterocyclic radical as defined above having one or morehalo, (lower)alkyl, (lower)alkoxy or phenyl substituents, Q is hydrogen,C₁ -C₁₀ alkyl, benzyl or phenethyl, and R" is hydrogen or an easilycleavable ester selected from the group consisting of benzhydryl,benzyl, p-nitrobenzyl, p-methoxybenzyl, trichloroethyl, trimethylsilyl,phenacyl, acetonyl, (lower)alkyl, triphenylmethyl, methoxymethyl,indanyl, phthalidyl, pivaloyloxymethyl and acetoxymethyl, or apharmaceutically acceptable salt thereof.
 2. A compound of claim 1wherein R is an acyl group of the formula

    R.sup.a C.sub.n H.sub.2n CO--

ps in which R^(a) is phenyl; phenyl substituted by one or more chloro,bromo, iodo, fluoro, nitro, amino, (lower)alkyl, guanidino,(lower)alkylthio, cyano, (lower)alkoxy, sulfamyl, (lower)alkylamino,hydroxy, acetoxy or trifluoromethyl radicals; 2-thienyl; 3-thienyl;tetrazolyl; sydnone-3- or sydnone-4; furyl; isothiazolyl; thiadiazolyloptionally substituted with phenyl; oxadiazolyl optionally substitutedwith phenyl; thiazolyl; imidazolyl; triazolyl; oxazolyl; pyridyl;furazan optionally substituted at the 3-position with methoxy;4-isoxazolyl optionally substituted at the 5-position with methyl and atthe 3-position with phenyl or halophenyl; 1,4-cyclohexadienyl;1-cyclohexenyl or 1-aminocyclohexyl.
 3. A compound of claim 2 wherein nis
 1. 4. A compound of claim 1 wherein R is an acyl group of the formula##STR293## in which R^(a) is phenyl and R^(b) and R^(c) which may be thesame or different each represent hydrogen, phenyl, benzyl, phenethyl orC₁ -C₆ alkyl.
 5. A compound of claim 1 and wherein R is an acyl group ofthe formula ##STR294## in which R^(a) is phenyl; phenyl substituted withone or more chloro, bromo, iodo, fluoro, nitro, amino, (lower)alkyl,(lower)alkylthio, cyano, (lower)alkoxy, (lower)alkylamino, hydroxy,acetoxy or trifluoromethyl radicals; 3-pyridyl or 4-pyridyl; and R^(b)and R^(c) are hydrogen.
 6. A compound of claim 1 wherein R is an acylgroup of the formula

    R.sup.a CO--

in which R^(a) is phenyl; phenyl substituted with one or more chloro,bromo, iodo, fluoro, nitro, amino, (lower)alkyl, (lower)alkylthio,cyano, (lower)alkoxy, (lower)alkylamino, di(lower)alkylamino, hydroxy,acetoxy or trifluoromethyl radicals; 2-ethoxynaphthoyl;3-phenyl-5-methylisoxaxol-4-yl; 3-o-chlorophenyl-5-methylisoxazol-4-yl;3-(2,6-dichlorophenyl)-5-methylisoxazol-4-yl; or 1-aminocyclohexyl.
 7. Acompound of claim 1 wherein R is an acyl group of the formula ##STR295##in which R^(a) is (a) aryl selected from the group consisting of phenyl,2-thienyl, 3-thienyl, furyl, 4-isoxazolyl, pyridyl, tetrazolyl,sydnone-3 or -4, imidazolyl, naphthoyl, quinoxalinyl, triazolyl,isothiazolyl, thiadiazolyl, thiazolyl, oxazolyl, oxadiazolyl, pyrazolyl,furazan, pyrazinyl, pyrimidinyl, pyridazinyl and triazinyl; (b)substituted aryl in which the aryl groups defined above under (a) aresubstituted by one or more chloro, bromo, iodo, fluoro, nitro, amino,cyano, (lower)alkanoyloxy, (lower)-alkanoyl, (lower)alkoxyamino,(lower)alkoxy, (lower)alkyl, (lower)alkylamino, hydroxy, guanidino,(lower)alkylthio, carboxy, phenyl, halophenyl, trifluoromethyl,di(lower)-alkylamino, sulfamyl, (lower)alkanoylamino,phenyl(lower)-alkylamido, cycloalkylamino, allylamido,morpholinocarbonyl, pyrrolidinocarbonyl, piperidinocarbonyl,tetrahydropyridino, furfurylamido or N-alkyl-N-anilino radicals; (c) C₃-C₁₂ cycloalkyl; (d) substituted C₃ -C₁₂ cycloalkyl in which thesubstituents are one or more chloro, bromo, fluoro, iodo, nitro,trifluoromethyl, C₁ -C₄ alkyl, C₁ -C₄ alkylamino, C₁ -C₂ alkoxy or aminoradicals; (e) C₃ -C₁₂ cycloalkenyl, said cycloalkenyl group having 1 or2 double bonds; or (f) substituted C₃ -C₁₂ cycloalkenyl, saidcycloalkenyl group having 1 or 2 double bonds and being substituted byone or more chloro, bromo, fluoro, iodo, nitro, trifluoromethyl, C₁ -C₄alkyl, C₁ -C₄ alkylamino, C₁ -C₂ alkoxy or amino radicals; and Y ishydrazino, guanidino, ureido; substituted ureido of the formula##STR296## in which R^(p) is hydrogen or C₁ -C₈ alkyl and R^(q) ishydrogen, C₁ -C₈ alkyl, C₂ -C₈ alkenyl, phenyl, benzoyl, C₁ -C₈ alkoxy-C₁ -C₈ alkyl or (carbo--C₁ -C₈ alkoxy)C₁ -C₈ alkyl; allophanamido;3-guanyl-1-ureido; 3-(2-furoyl)ureido; 3-(benzoyl)ureido; cyano;cyanamino; azido; amino; a group obtained by reacting the amino group Ywith acetone, formaldehyde, acetaldehyde, butyraldehyde, acetylacetone,methyl acetoacetate, benzaldehyde, salicylaldehyde, methyl ethyl ketoneor ethyl acetoacetate; hydroxy; (lower)alkoxy; carboxy;5-indanyloxycarbonyl; triazolyl; tetrazolyl; halogeno; formyloxy;(lower)alkanoyloxy; sulfo; or sulfoamino.
 8. A compound of claim 1wherein R is an acyl group of the formula ##STR297## in which R^(a) is2-thienyl; 3-thienyl; phenyl; or phenyl substituted by one or morenitro, di(lower)alkylamino, (lower)alkanoylamino, amino, hydroxy,(lower)alkanoyloxy, C₁ -C₆ alkyl, C₁ -C₆ alkoxy, sulfamyl, chloro,bromo, iodo, fluoro or trifluoromethyl radicals; X is oxygen; X' isoxygen or imino; and R^(i) is (lower)alkyl, phenyl, 2-thienyl,3-thienyl, 2-furyl or 5-nitro-2-furyl.
 9. A compound of claim 8 whereinR^(a) is phenyl, p-hydroxyphenyl, 2-thienyl or 3-thienyl; X' is oxygen;and R^(i) is phenyl or 2-furyl.
 10. A compound of claim 1 wherein R isan acyl group of the formula ##STR298## in which Ar' is a radical of theformula ##STR299## in which R^(m), R^(n) and R^(o) are alike ordifferent and each is hydrogen, hydroxy (lower)alkyl, cyano,(lower)alkoxy, chloro, bromo, iodo, fluoro, trifluoromethyl, nitro,amino, (lower)alkylamino, di(lower)-alkylamino, (lower)alkanoyl,(lower)alkanoyloxy or phenyl and Y is amino or a group obtained byreacting the amino group with acetaldehyde, formaldehyde or acetone;fluoro; chloro; bromo; iodo; hydroxy; (lower)alkanoyloxy; carboxy;guanidino; 3-guanyl-1-ureido; 3-(2-furoyl)ureido; 3-benzoylureido;sulfo; sulfoamino; ureido; thioureido; (lower)alkoxy; cyano; cyanamino;or indanyloxycarbonyl.
 11. A compound of claim 10 wherein Ar' is phenyl,p-hydroxyphenyl, 4-hydroxy-3,5-dichlorophenyl, 3-chloro-4-hydroxyphenyl,o-, m- or p-aminomethylphenyl, 2-thienyl, 3-thienyl, 1-cyclohexenyl or1,4-cyclohexadienyl and Y is amino, hydroxy or carboxy.
 12. A compoundof claim 1 wherein R is an acyl group of the formula ##STR300##
 13. Acompound of claim 1 wherein R is an acyl group of the formula ##STR301##wherein U and V are alike or different and each is hydrogen, chloro orfluoro; ##STR302##
 14. An acid of claim 1 wherein R" is hydrogen and Ris phenoxyacetyl, or a pharmaceutically acceptable salt thereof.
 15. Anacid of claim 1 wherein R" is hydrogen and R is phenylacetyl, or apharmaceutically acceptable salt thereof.
 16. An acid of claim 1 whereinR" is hydrogen and R is α-aminophenylacetyl, or a pharmaceuticallyacceptable salt thereof.
 17. The D-isomer of the acid of claim 16 or apharmaceutically acceptable salt thereof.
 18. An acid of claim 1 whereinR" is hydrogen and R is α-carboxyphenylacetyl, cyanoacetyl,α-amino-α-(p-hydroxyphenyl)-acetyl,α-amino-α-(3-chloro-4-hydroxyphenyl)acetyl,α-amino-α-(3,5-dichloro-4-hydroxyphenyl)acetyl,α-amino-α-(2-thienyl)acetyl, α-amino-α-(3-thienyl)acetyl,α-amino-α-(1-cyclohexenyl)acetyl, α-amino-α-(1,4-cyclo-hexadienyl)acetyl, α-hydroxyacetyl, α-hydroxy-α-(2-thienyl)acetyl,α-hydroxy-α-(3-thienyl)acetyl, α-hydroxy-α-(1-cyclohexenyl)acetyl,α-hydroxy-α-(1,4-cyclohexadienyl)acetyl, α-carboxy-α-(2-thienyl)acetyl,α-carboxy-α-(3-thienyl)acetyl, α-carboxy-α-(1-cyclohexenyl)acetyl,α-carboxy-α-(1,4-cyclohexadienyl)acetyl,α-indanyloxycarbonyl-α-phenylacetyl, 1-(1H)-tetrazolyl,4-pyridylthioacetyl, 2-thienylacetyl, 3-thienylacetyl,1-cyclohexenylacetyl, 1,4-cyclohexadienylacetyl,O-aminomethylphenylacetyl, 1-aminocyclohexylcarbonyl,2,6-dimethoxybenzoyl, sydnoneacetyl or α-azidophenylacetyl, or apharmaceutically acceptable salt thereof.
 19. The D-isomer of an acid ofclaim 1 wherein R" is hydrogen and R isα-amino-α-(p-hydroxyphenyl)acetyl,α-amino-α-(3-chloro-4-hydroxyphenyl)acetyl,α-amino-α-(3,5-dichloro-4-hydroxyphenyl)acetyl,α-amino-α-(2-thienyl)acetyl, α-amino-α-(3-thienyl)acetyl,α-amino-α-(1-cyclohexenyl)acetyl, α-amino-α-(1,4-cyclohexadienyl)acetyl,α-hydroxyacetyl, α-hydroxy-α-(2-thienyl)acetyl,α-hydroxy-α-(3-thienyl)acetyl, α-hydroxy-α-(1-cyclohexenyl)acetyl orα-hydroxy-α-(1,4-cyclohexadienyl)acetyl, or a pharmaceuticallyacceptable salt thereof.
 20. A compound of the formula ##STR303##wherein R represents an acyl group of the formula

    R.sup.a C.sub.n H.sub.2n CO--                              (i)

wherein R^(a) is (a) aryl selected from the group consisting of phenyl,2-thienyl, 3-thienyl, furyl, 4-isoxazolyl, pyridyl, tetrazolyl,sydnone-3 or -4, imidazolyl, napthoyl, quinoxalinyl, triazolyl,isothiazolyl, thiadiazolyl, thiazolyl, oxazolyl, oxadiazolyl, pyrazolyl,furazan, pyrazinyl, pyrimidinyl, pyridazinyl and triazinyl; (b)substituted aryl in which the aryl groups define above under (a) aresubstituted by one or more chloro, bromo, iodo, fluoro, nitro, amino,cyano, (lower)alkanoyloxy, (lower)alkanoyl, (lower)alkoxyamino,(lower)alkoxy, (lower)alkyl, (lower)alkylamino, hydroxy, guanidino,(lower)alkylthio, carboxy, phenyl, halophenyl, trifluoromethyl,di(lower)alkylamino, sulfamyl, (lower)alkanoylamino,phenyl(lower)alkylamido, cycloalkylamino, allylamido,morpholinocarboxyl, pyrrolidinocarbonyl, piperidinocarbonyl,tetrahydropyridino, furfurylamido or N-alkyl-N-anilino radicals; (c) C₃-C₁₂ cycloalkyl; (d) substituted C₃ -C₁₂ cycloalkyl in which thesubstitutents are one or more chloro, bromo, fluoro, iodo, nitro,trifluoromethyl, C₁ -C₄ alkyl, C₁ -C₄ alkylamino, C₁ -C₂ alkoxy or aminoradicals; (e) C₃ -C₁₂ cycloalkenyl, said cycloalkenyl group having 1 or2 double bonds; or (f) substituted C₃ -C₁₂ cycloalkenyl, saidcycloalkenyl group having 1 or 2 double bonds and being substituted byone or more chloro, bromo, fluoro, iodo, nitro, trifluoromethyl, C₁ -C₄alkyl, C₁ -C₄ alkylamino, C₁ -C₂ alkoxy or amino radicals; and n is aninteger from 1-4;

    C.sub.n H.sub.2n+1 CO--                                    (ii)

wherein n is an integer from 1-7, the alkyl portion of said acyl groupbeing straight or branched and optionally interrupted by an oxygen orsulfur atom;

    C.sub.n H.sub.2n-1 CO--                                    (iii)

wherein n is an integer from 2-7, the alkenyl portion of said acyl groupbeing straight or branched and optionally interrupted by an oxygen orsulfur atom; ##STR304## wherein R^(a) is as defined above under (i) andin addition may be benzyl, C₁ -C₆ alkyl or (lower)alkoxy carbonyl andR^(b) and R^(c) which may be the same or different each representhydrogen, phenyl, benzyl, phenethyl or C₁ -C₆ alkyl; ##STR305## whereinR^(a) is as defined above under (i) and in addition may be benzyl or C₁-C₆ alkyl and R^(b) and R^(c) are as defined under (iv);

    R.sup.a X(CH.sub.2).sub.m CO--                             (vi)

wherein R^(a) is as defined under (i) and in addition may be benzyl; Xis oxygen or sulfur; and m is an integer of 2-5;

    R.sup.a CO--                                               (vii)

wherein R^(a) is as defined under (i); ##STR306## wherein R^(a) is asdefined under (i) and Y is hydrazino, guanidino, ureido; substitutedureido of the formula ##STR307## in which R^(p) is hydrogen or C₁ -C₈alkyl and R^(q) is hydrogen, C₁ -C₈ alkyl, C₂ -C₈ alkenyl, phenyl,benzoyl, C₁ -C₈ alkoxy-C₁ -C₈ alkyl or (carbo-C₁ -C₈ alkoxy)C₁ -C₈alkyl; allophanamido; 3-guanyl-1-ureido; 3-(2-furoyl)ureido;3-(benzoyl)ureido; cyano; cyanamino; azido; amino; a group obtained byreacting the amino group Y with acetone, formaldehyde, acetaldehyde,butyraldehyde, acetylacetone, methyl acetoacetate, benzaldehyde,salicylaldehyde, methyl ethyl ketone or ethyl acetoacetate; hydroxy;(lower)alkoxy; carboxy; 5-indanyloxycarbonyl; triazolyl; tetrazolyl;halogeno; formyloxy; (lower)alkanoyloxy; sulfo; or sulfoamino;##STR308## wherein R^(d), R^(e) and R^(f) which may be the same ordifferent may each represent C₁ -C₆ alkyl, phenyl or phenyl substitutedby one or more chloro, bromo, iodo, fluoro, trifluoromethyl, nitro,amino, cyano, (lower)alkanoyloxy, (lower)alkanoyl, (lower)alkoxyamino,(lower)alkoxy, (lower)alkyl, (lower)alkylamino, hydroxy,(lower)alkylthio, carboxy, di(lower)alkylamino or sulfamyl radicals;##STR309## wherein R^(a) is as defined under (i) and in addition may behydrogen, C₁ -C₆ alkyl, halogen-substituted C₁ -C₆ alkyl, phenethyl,phenoxymethyl, benzyl or R^(a) -CO- and X is oxygen or sulfur;##STR310## wherein Y is as defined under (viii) and n is an integer of1-4;

    R.sup.g CH(NH.sub.2) (CH.sub.2).sub.n CO--                 (xii)

wherein n is an integer of 1-10, or

    H.sub.2 N--C.sub.n H.sub.2n Ar(CH.sub.2).sub.m CO--

wherein m is 0 or an integer from 1-10, and n is 0, 1 or 2; R^(g) ishydrogen, (lower)alkyl, phenyl, benzyl or carboxy and Ar is p-phenyleneor 1,4-naphthylene;

    R.sup.h CO-CO--                                            (xiii)

wherein R^(h) is 2-thienyl; 3-thienyl; α-naphthyl; 2-phenanthryl or amono-, di- or tri-substituted phenyl group, the substituents beingchloro, bromo, iodo, fluoro, amino, di(lower)alkylamino, (lower)alkyl,(lower)alkoxy, nitro or (lower)alkanoylamino; ##STR311## wherein R^(a)is as defined under (i); X is oxygen or sulfur; X' is oxygen or imino;and R^(i) is (lower)alkyl, cycloalkyl having 4, 5, 6 or 7 carbon atoms,monohalo (lower)alkyl, dichloromethyl, trichloromethyl, (lower)alkenylof 2-6 carbon atoms, ##STR312## n is an integer from 0 to 3 inclusiveand each of R^(k) and R^(j) is hydrogen, nitro, di(lower)alkylamino,(lower)alkanoylamino, (lower)alkanoyloxy, C₁ -C₆ alkyl, C₁ -C₆ alkoxy,sulfamyl, chloro, iodo, bromo, fluoro, or trifluoromethyl; ##STR313##wherein R^(a) is as defined under (i) and R^(i) is as defined under(xiv); or ##STR314## wherein R^(a) is as defined under (i) and R¹ is(lower)alkyl, cycloalkyl of 3-12 carbon atoms, phenyl, a monocyclicheterocyclic radical having 5 or 6 atoms exclusive of hydrogen which areC, S, N or O, no more than 2 atoms being other then C, or a substitutedmonocyclic heterocyclic radical as defined above having one or morehalo, (lower)alkyl, (lower)alkoxy or phenyl substituents, and R" ishydrogen or an easily cleavable ester selected from the group consistingof benzhydryl, benzyl, p-nitrobenzyl, p-methoxybenzyl, trichloroethyl,trimethylsilyl, phenacyl, acetonyl, (lower)alkyl, triphenylmethyl,methoxymethyl, indanyl, phthalidyl, pivaloyloxymethyl and acetoxymethyl,or a pharmaceutically acceptable salt thereof.
 21. A compound of claim20 wherein R is an acyl group of the formula

    R.sup.a C.sub.n H.sub.2n CO--

in which R^(a) is phenyl; phenyl substituted by one or more chloro,bromo, iodo, fluoro, nitro, amino, (lower)alkyl, guanidino,(lower)alkylthio, cyano, (lower)alkoxy, sulfamyl, (lower)alkylamino,hydroxy, acetoxy or trifluoromethyl radicals; 2-thienyl; 3-thienyl;tetrazolyl; sydnone-3- or sydnone-4; furyl; isothiazolyl; thiadiazolyloptionally substituted with phenyl; oxadiazolyl optionally substitutedwith phenyl; oxadiazolyl optionally substituted with phenyl; thiazolyl;imidazolyl; triazolyl; oxazolyl; pyridyl; furazan optionally substitutedat the 3-position with methoxy, 4-isoxazolyl optionally substituted atthe 5-position with methyl and at the 3-position with phenyl orhalophenyl; 1,4-cyclohexadienyl; 1-cyclohexenyl or 1-aminocyclohexyl.22. A compound of claim 21 wherein n is
 1. 23. A compound of claim 20wherein R is an acyl group of the formula ##STR315## in which R^(a) isphenyl and R^(b) and R^(c) which may be the same or different eachrepresent hydrogen, phenyl, benzyl, phenethyl or C₁ -C₆ alkyl.
 24. Acompound of claim 20 wherein R is an acyl group of the formula##STR316## in which R^(a) is phenyl; phenyl substituted with one or morechloro, bromo, iodo, fluoro, nitro, amino, (lower)alkyl,(lower)alkylthio, cyano, (lower)alkoxy, (lower)alkylamino, hydroxy,acetoxy or trifluoromethyl radicals; 3-pyridyl or 4-pyridyl; and R^(b)and R^(c) are hydrogen.
 25. A compound of claim 20 wherein R is an acylgroup of the formula

    R.sup.a CO--

in which R^(a) is phenyl; phenyl substituted with one or more chloro,bromo, iodo, fluoro, nitro, amino, (lower)alkyl, (lower)alkylthio,cyano, (lower)alkoxy, (lower)alkylamino, di(lower)alkylamino, hydroxy,acetoxy or trifluoromethyl radicals; 2-ethoxynaphthoyl;3-phenyl-5-methylisoxazol-4-yl; 3-o-chlorophenyl-5-methylisoxazol-4-yl;3-(2,6-dichlorophenyl)-5-methylisoxazol-4-yl; or 1-aminocyclohexyl. 26.A compound of claim 20 wherein R is an acyl group of the formula##STR317## in which R^(a) is (a) aryl selected from the group consistingof phenyl, 2-thienyl, 3-thienyl, furyl, 4-isoxazolyl, pyridyl,tetrazolyl, sydnone-3 or -4, imidazolyl, naphthoyl, quinoxalinyl,triazolyl, isothiazolyl, thiadiazolyl, thiazolyl, oxazolyl, oxadiazolyl,pyrazolyl, furazan, pyrazinyl, pyrimidinyl, pyridazinyl and triazinyl;(b) substituted aryl in which the aryl groups defined above under (a)are substituted by one or more chloro, bromo, iodo, fluoro, nitro,amino, cyano, (lower)alkanoyloxy, (lower)alkanoyl, (lower)alkoxyamino,(lower)alkoxy, (lower)alkyl, (lower)alkylamino, hydroxy, guanidino,(lower)alkylthio, carboxy, phenyl, halophenyl, trifluoromethyl,di(lower)alkylamino, sulfamyl, (lower)alkanoylamino,phenyl(lower)alkylamido, cycloalkylamino, allylamido,morpholinocarbonyl, pyrrolidinocarbonyl, piperidinocarbonyl,tetrahydropyridino, furfurylamido or N-alkyl-N-anilino radicals; (c) C₃-C₁₂ cycloalkyl; (d) substituted C₃ -C₁₂ cycloalkyl in which thesubstituents are one or more chloro, bromo, fluoro, iodo, nitro,trifluoromethyl, C₁ -C₄ alkyl, C₁ -C₄ alkylamino, C₁ -C₂ alkoxy or aminoradicals; (e) C₃ -C₁₂ cycloalkenyl, said cycloalkenyl group having 1 or2 double bonds; or (f) substituted C₃ -C₁₂ cycloalkenyl, saidcycloalkenyl group having 1 or 2 double bonds and being substituted byone or more chloro, bromo, fluoro, iodo, nitro, trifluoromethyl, C₁ -C₄alkyl, C₁ -C₄ alkylamino, C₁ -C₂ alkoxy or amino radicals; and Y ishydrazino, guanidino, ureido; substituted ureido of the formula##STR318## in which R^(p) is hydrogen or C₁ -C₈ alkyl and R^(q) ishydrogen, C₁ -C₈ alkyl, C₂ -C₈ alkenyl, phenyl, benzoyl, C₁ -C₈ alkoxyC₁ -C₈ alkyl or (carbo--C₁ -C₈ alkoxy)C₁ -C₈ alkyl; allophanamido;3-guanyl-1-ureido; 3-(2-furoyl)ureido; 3-(benzoyl)ureido; cyano;cyanamino; azido; amino; a group obtained by reacting the amino group Ywith acetone, formaldehyde, acetaldehyde, butyraldehyde, acetylacetone,methyl acetoacetate, benzaldehyde, salicylaldehyde, methyl ethyl ketoneor ethyl acetoacetate; hydroxy; (lower)alkoxy; carboxy;5-indanyloxycarbonyl; triazolyl; tetrazolyl; halogeno; formyloxy;(lower)alkanoyloxy; sulfo; and sulfoamino.
 27. A compound of claim 20wherein R is an acyl group of the formula ##STR319## in which R^(a) is2-thienyl; 3-thienyl; phenyl; or phenyl substituted by one or morenitro, di(lower)alkylamino, (lower)alkanoylamino, amino, hydroxy,(lower)alkanoyloxy, C₁ -C₆ alkyl; C₁ -C₆ alkoxy, sulfamyl, chloro,bromo, iodo, fluoro or trifluoromethyl radicals; X is oxygen; X' isoxygen or imino; and R^(i) is (lower)alkyl, phenyl, 2-thienyl,3-thienyl, 2-furyl or 5-nitro-2-furyl.
 28. A compound of claim 27wherein R^(a) is phenyl, p-hydroxyphenyl, 2-thienyl or 3-thienyl; X' isoxygen; and R^(i) is phenyl or 2-furyl.
 29. A compound of claim 20wherein R is an acyl group of the formula ##STR320## in which Ar' is aradical of the formula ##STR321## in which R^(m), R^(n) and R^(o) arealike or different and each is hydrogen, hydroxy, (lower)alkyl, cyano,(lower)alkoxy, chloro, bromo, iodo, fluoro, trifluoromethyl, nitro,amino, (lower)alkylamino, di(lower)alkylamino, (lower)alkanoyl,(lower)alkanoyloxy or phenyl and Y is amino or a group obtained byreacting the amino group with acetaldehyde, formaldehyde or acetone;fluoro; chloro; bromo; iodo; hydroxy; (lower)alkanoyloxy; carboxy;guanidino; 3-guanyl-1-ureido; 3-(2-furoyl)ureido; 3-benzoylureido;sulfo; sulfoamino; ureido; thioureido; (lower) alkoxy; cyano, cyanamino;or indanyloxycarbonyl.
 30. A compound of claim 29 wherein Ar' is phenyl,p-hydroxyphenyl, 4-hydroxy-3,5-dichlorophenyl, 3-chloro-4-hydroxyphenyl,o-, m- or p-aminomethylphenyl, 2-thienyl, 3-thienyl, 1-cyclohexenyl or1,4-cyclohexadienyl and Y is amino, hydroxy or carboxy.
 31. A compoundof claim 20 wherein R is an acyl group of the formula ##STR322##
 32. Acompound of claim 20 wherein R is an acyl group of the formula##STR323## wherein U and V are alike or different and each is hydrogen,chloro or fluoro; ##STR324##
 33. An acid of claim 20 wherein R" ishydrogen and R is phenoxyacetyl, or a pharmaceutically acceptable saltthereof.
 34. An acid of claim 20 wherein R" is hydrogen and R isphenylacetyl, or a pharmaceutically acceptable salt thereof.
 35. An acidof claim 20 wherein R" is hydrogen and R is α-aminophenylacetyl, or apharmaceutically acceptable salt thereof.
 36. The D-isomer of the acidof claim 35, or a pharmaceutically acceptable salt thereof.
 37. Thepivaloyloxymethyl, methoxymethyl, phthalidyl, indanyl or acetoxymethylester of the acid of claim 35, or a pharmaceutically acceptable saltthereof.
 38. An acid of claim 20 wherein R" is hydrogen and R isα-carboxyphenylacetyl, cyanoacetyl, α-amino-α-(p-hydroxyphenyl)acetyl,α-amino-α-(3-chloro-4-hydroxyphenyl)acetyl,α-amino-α-(3,5-dichloro-4-hydroxyphenyl)acetyl,α-amino-α-(2-thienyl)acetyl, α-amino-α-(3-thienyl)acetyl,α-amino-α-(1-cyclohexenyl)acetyl, α-amino-α-(1,4-cyclohexadienyl)acetyl,α-hydroxyacetyl, α-hydroxy-α-(2-thienyl)acetyl,α-hydroxy-α-(3-thienyl)acetyl, α-hydroxy-α-(1-cyclohexenyl)acetyl,α-hydroxy-α-(1,4-cyclohexadienyl)acetyl, α-carboxy-α-(2-thienyl)acetyl,α-carboxy-α-(3-thienyl)acetyl, α-carboxy-α-(1-cyclohexenyl)acetyl,α-carboxy-α-(1,4-cyclohexadienyl)-acetyl,α-indanyloxycarbonyl-α-phenylacetyl, 1-(1H)-tetrazolyl,4-pyridylthioacetyl, 2-thienylacetyl, 3-thienylacetyl,1-cyclohexenylacetyl, 1,4-cyclohexadienylacetyl,o-aminomethylphenylacetyl, 1-aminocyclohexylcarbonyl,2,6-dimethoxybenzoyl, sydnoneacetyl or α-azidophenylacetyl, or apharmaceutically acceptable salt thereof.
 39. The pivaloyloxymethyl,methoxymethyl, phthalidyl, indanyl or acetoxymethyl ester of an acid ofclaim 38, or a pharmaceutically acceptable salt thereof.
 40. TheD-isomer of an acid of claim 20 wherein R" is hydrogen and R isα-amino-α-(p-hydroxyphenyl)acetyl,α-amino-α-(3-chloro-4-hydroxyphenyl)acetyl,α-amino-α-(3,5-dichloro-4-hydroxyphenyl)acetyl,α-amino-α-(2-thienyl)acetyl, α-amino-α-(3-thienyl)acetyl,α-amino-α-(1-cyclohexenyl)acetyl, α-amino-α-(1,4-cyclohexadienyl)acetyl,α-hydroxyacetyl, α-hydroxy-α-(2-thienyl)acetyl,α-hydroxy-α-(3-thienyl)acetyl, α-hydroxy-α-(1-cyclohexenyl)acetyl orα-hydroxy-α-(1,4-cyclohexadienyl)acetyl, or a pharmaceuticallyacceptable salt thereof.
 41. The pivaloyloxymethyl, acetoxymethyl,phthalidyl, indanyl or methoxymethyl ester of an acid of claim 40, or apharmaceutically acceptable salt thereof.
 42. An acid of claim 20wherein R" is hydrogen and R is ##STR325## or a pharmaceuticallyacceptable salt thereof.
 43. An acid of claim 20 wherein R" is hydrogenand R is α-carboxyphenylacetyl, or a pharmaceutically acceptable saltthereof.
 44. An acid of claim 20 wherein R" is hydrogen and R iso-hydroxyphenylacetyl, or a pharmaceutically acceptable salt thereof.45. An acid of claim 20 wherein R" is hydrogen and R is cyanoacetyl, ora pharmaceutically acceptable salt thereof.
 46. An acid of claim 20wherein R" is hydrogen and R is ##STR326## or a pharmaceuticallyacceptable salt thereof.
 47. An acid of claim 20 wherein R" is hydrogenand R is CH₃ CH₂ OCO-, or a pharmaceutically acceptable salt thereof.48. The D-isomer of an acid of claim 20 wherein R" is hydrogen and R isα-amino-α-(p-hydroxyphenyl)acetyl, or a pharmaceutically acceptable saltthereof.
 49. The D-isomer of an acid of claim 20 wherein R" is hydrogenand R is α-amino-α-(3-chloro-4-hydroxyphenyl)acetyl, or apharmaceutically acceptable salt thereof.
 50. The D-isomer of an acid ofclaim 20 wherein R" is hydrogen and R isα-amino-α-(3,5-dichloro-4-hydroxyphenyl)acetyl, or a pharmaceuticallyacceptable salt thereof.
 51. The D-isomer of an acid of claim 20 whereinR" is hydrogen R is α-amino-α-(2-thienyl)acetyl, or a pharmaceuticallyacceptable salt thereof.
 52. The D-isomer of an acid of claim 20 whereinR" is hydrogen and R is α-amino-α-(3-thienyl)acetyl, or apharmaceutically acceptable salt thereof.
 53. The D-isomer of an acid ofclaim 20 wherein R" is hydrogen and R is ##STR327## or apharmaceutically acceptable salt thereof.
 54. The D-isomer of an acid ofclaim 20 wherein R" is hydrogen and R is ##STR328## or apharmaceutically acceptable salt thereof.
 55. The D-isomer of an acid ofclaim 20 wherein R" is hydrogen and R is α-hydroxy-α-(2-thienyl)acetyl,or a pharmaceutically acceptable salt thereof.
 56. The D-isomer of anacid of claim 20 wherein R" is hydrogen and R isα-hydroxy-α-(3-thienyl)acetyl, or a pharmaceutically acceptable saltthereof.
 57. An acid of claim 20 wherein R" is hydrogen and R isα-carboxy-α-(2-thienyl)acetyl, or a pharmaceutically acceptable saltthereof.
 58. An acid of claim 20 wherein R" is hydrogen and R isα-carboxy-α-(3-thienyl)acetyl, or a pharmaceutically acceptable saltthereof.
 59. An acid of claim 20 wherein R" is hydrogen and R iso-aminomethylphenylacetyl, or a pharmaceutically acceptable saltthereof.
 60. An acid of claim 20 wherein R" is hydrogen and R iso-aminomethylphenylpropionyl, or a pharmaceutically acceptable saltthereof.
 61. An acid of claim 20 wherein R" is hydrogen and R is##STR329## or a pharmaceutically acceptable salt thereof.
 62. An acid ofclaim 20 wherein R" is hydrogen and R is ##STR330## or apharmaceutically acceptable salt thereof.
 63. An acid of claim 20wherein R" is hydrogen and R is ##STR331## or a pharmaceuticallyacceptable salt thereof.
 64. An acid of claim 20 wherein R" is hydrogenand R is 2-thienylacetyl, or a pharmaceutically acceptable salt thereof.65. An acid of claim 20 wherein R" is hydrogen and R is 3-thienylacetyl,or a pharmaceutically acceptable salt thereof.
 66. An acid of claim 20wherein R" is hydrogen and R is ##STR332## or a pharmaceuticallyacceptable salt thereof.
 67. An acid of claim 20 wherein R" is hydrogenand R is ##STR333## or a pharmaceutically acceptable salt thereof. 68.An acid of claim 22 wherein R" is hydrogen and R is ##STR334## or apharmaceutically acceptable salt thereof.
 69. An acid of claim 20wherein R" is hydrogen and R is ##STR335## or a pharmaceuticallyacceptable salt thereof.
 70. An acid of claim 20 wherein R" is hydrogenand R is

    CH.sub.3 (CH.sub.2).sub.3 CO--,

or a pharmaceutically acceptable salt thereof.
 71. An acid of claim 20wherein R" is hydrogen and R is ##STR336## or a pharmaceuticallyacceptable salt thereof.
 72. An acid of claim 20 wherein R" is hydrogenand R is ##STR337## or a pharmaceutically acceptable salt thereof. 73.An acid of claim 20 wherein R" is hydrogen and R is ##STR338## or apharmaceutically acceptable salt thereof.
 74. An acid of claim 20wherein R" is hydrogen and R is ##STR339## or a pharmaceuticallyacceptable salt thereof.
 75. An acid of claim 20 wherein R" is hydrogenand R is ##STR340## or a pharmaceutically acceptable salt thereof. 76.An acid of claim 20 wherein R" is hydrogen and R is ##STR341## or apharmaceutically acceptable salt thereof
 77. An acid of claim 20 whereinR" is hydrogen and R is ##STR342## or a pharmaceutically acceptable saltthereof.
 78. An acid of claim 20 wherein R" is hydrogen and R is##STR343## or a pharmaceutically acceptable salt thereof.